LOCUS Exported 7166 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of ChrimsonR-tdTomato under the Syn promoter, in floxed/forward (Cre-dependent) manner. Cre turns gene off. Using bGHpA signal. tdTomato is a codon diversified version. . ACCESSION . VERSION . KEYWORDS pAAV-Syn-FLEX-fd [ChrimsonR-tdTomato] SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7166) AUTHORS Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK, Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, Constantine-Paton M, Wong GK, Boyden ES TITLE Independent optical excitation of distinct neural populations. JOURNAL Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 2014 Feb 9. PUBMED 24509633 REFERENCE 2 (bases 1 to 7166) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7166 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter rep_origin complement(1932..2387) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2069..2090) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2281..2300 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2459..2588 /label=AAV2 ITR promoter 2691..3138 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind complement(3154..3187) /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(3250..3283) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 3294..3303 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3300..4349 /codon_start=1 /product="red light-drivable channelrhodopsin from Chlamydomonas noctigama (Klapoetke et al., 2014)" /label=ChrimsonR /note="K176R mutant with accelerated closing kinetics" /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE FVEEEDEDTV" protein_bind 5806..5839 /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 5910..5943 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5976..6564 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6029..6049) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6447..6458) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6605..6812 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6982..7111 /label=AAV2 ITR ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag 2641 gaagatcgta ggttaattaa gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg 2701 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca 2761 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg 2821 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct 2881 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca 2941 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc 3001 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg 3061 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg 3121 tcgtgcctga gagcgcagtc gagaagaatt cccataactt cgtataaagt atcctatacg 3181 aagttatatc aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac 3241 catcgaccca taacttcgta taatgtatgc tatacgaagt tatactagct agcgccacca 3301 tggctgagct gatcagcagc gccaccagat ctctgtttgc cgccggaggc atcaaccctt 3361 ggcctaaccc ctaccaccac gaggacatgg gctgtggagg aatgacacct acaggcgagt 3421 gcttcagcac cgagtggtgg tgtgaccctt cttacggact gagcgacgcc ggatacggat 3481 attgcttcgt ggaggccaca ggcggctacc tggtcgtggg agtggagaag aagcaggctt 3541 ggctgcacag cagaggcaca ccaggagaaa agatcggcgc ccaggtctgc cagtggattg 3601 ctttcagcat cgccatcgcc ctgctgacat tctacggctt cagcgcctgg aaggccactt 3661 gcggttggga ggaggtctac gtctgttgcg tcgaggtgct gttcgtgacc ctggagatct 3721 tcaaggagtt cagcagcccc gccacagtgt acctgtctac cggcaaccac gcctattgcc 3781 tgcgctactt cgagtggctg ctgtcttgcc ccgtgatcct gatcagactg agcaacctga 3841 gcggcctgaa gaacgactac agcaagcgga ccatgggcct gatcgtgtct tgcgtgggaa 3901 tgatcgtgtt cggcatggcc gcaggactgg ctaccgattg gctcaagtgg ctgctgtata 3961 tcgtgtcttg catctacggc ggctacatgt acttccaggc cgccaagtgc tacgtggaag 4021 ccaaccacag cgtgcctaaa ggccattgcc gcatggtcgt gaagctgatg gcctacgctt 4081 acttcgcctc ttggggcagc tacccaatcc tctgggcagt gggaccagaa ggactgctga 4141 agctgagccc ttacgccaac agcatcggcc acagcatctg cgacatcatc gccaaggagt 4201 tttggacctt cctggcccac cacctgagga tcaagatcca cgagcacatc ctgatccacg 4261 gcgacatccg gaagaccacc aagatggaga tcggaggcga ggaggtggaa gtggaagagt 4321 tcgtggagga ggaggacgag gacacagtgg cggcaccggt agtagcagtg agtaagggcg 4381 aggaagtgat caaagagttc atgcggttta aggtgagaat ggaaggaagc atgaacggcc 4441 acgagttcga aattgaggga gaaggagagg gacggcccta cgagggcacc cagacagcca 4501 agctgaaagt gacaaagggc gggcctctgc cattcgcttg ggacatcctg agcccacagt 4561 ttatgtacgg ctccaaggcc tatgtgaaac atccagctga cattcccgat tataagaaac 4621 tgagcttccc cgaggggttt aagtgggaaa gagtgatgaa cttcgaggac ggaggcctgg 4681 tgactgtgac ccaggacagc tccctgcagg atgggaccct gatctacaag gtgaaaatga 4741 gagggacaaa ttttccccct gatggacctg tgatgcagaa gaaaactatg ggatgggagg 4801 cctccaccga aaggctgtat ccacgcgacg gggtgctgaa aggagaaatc caccaggctc 4861 tgaagctgaa agatggggga cattacctgg tggagttcaa gacaatctac atggccaaga 4921 aacctgtgca gctgccaggc tactattacg tggacacaaa actggatatc acttcacaca 4981 acgaggacta cactattgtg gagcagtatg aacggagcga ggggagacac catctgttcc 5041 tgggccatgg gactggaagt accggctcag ggtctagtgg aaccgcctca agcgaggata 5101 acaatatggc tgtgatcaaa gagttcatga ggtttaaggt gcgcatggag ggcagcatga 5161 atgggcacga atttgagatt gaaggagagg gcgaagggag gccttacgag ggcacacaga 5221 ctgccaagct gaaagtgacc aagggaggac cactgccttt cgcttgggat atcctgtctc 5281 ctcagtttat gtacggaagt aaggcctatg tcaagcatcc cgctgacatt cctgattaca 5341 agaaactgtc tttcccagag ggctttaagt gggagagagt gatgaatttt gaagatggag 5401 gcctggtgac cgtgacacag gactcctctc tgcaggatgg cactctgatc tacaaagtca 5461 aaatgcgcgg caccaatttt ccacccgatg ggcccgtgat gcagaagaaa acaatggggt 5521 gggaggccag cactgaacgg ctgtatccta gagacggagt gctgaagggc gaaatccacc 5581 aggccctgaa gctgaaagac ggcggccact acctggtgga gttcaaaacc atctacatgg 5641 ccaagaaacc agtgcagctg cccggctatt actatgtgga caccaagctg gatatcacat 5701 cccacaatga agactacacc attgtggaac agtatgagag gtctgaagga cgccaccatc 5761 tgtttctgta cggcatggat gagctgtata agtaaggtac ctagtataac ttcgtatagg 5821 atactttata cgaagttatc attgggattc ttcctatttt gatccaagca tcaccatcga 5881 ccctctagtc cagatctcac catcgaccca taacttcgta tagcatacat tatacgaagt 5941 tatgtccctc gaagaggttc gcgaagctta tcgataatca acctctggat tacaaaattt 6001 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 6061 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 6121 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 6181 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 6241 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 6301 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 6361 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 6421 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 6481 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 6541 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg ctcataaagc 6601 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 6661 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 6721 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 6781 gattgggaag acaatagcag gcatgctggg gatgcggtcc ggactgtaca attgcattca 6841 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta 6901 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg 6961 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc 7021 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc 7081 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg 7141 gagaggcggt ttgcgtattg ggcgct //