LOCUS Exported 5915 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Can be used to generate AAV virus that will express tdTomato in the presence of Cre in neurons from the synapsin promoter. ACCESSION . VERSION . KEYWORDS AAV phSyn1(S)-FLEX-tdTomato-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5915) AUTHORS Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C, Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA, Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E, Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW, Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones AR, Zeng H TITLE A mesoscale connectome of the mouse brain. JOURNAL Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub 2014 Apr 2. PUBMED 24695228 REFERENCE 2 (bases 1 to 5915) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5915 /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 177..624 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 657..690 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 701..734 /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 complement(1563..1586) /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(1761..1780) /label=mCherry-F /note="mCherry, forward primer" primer_bind 2030..2048 /label=mCherry-R /note="mCherry, reverse primer" regulatory 2178..2187 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2218..2251) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(2262..2295) /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)." misc_feature 2319..2907 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2372..2392) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2790..2801) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2942..2959) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2948..3155 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3178..3318 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3178..3307 /label=AAV2 ITR rep_origin 3393..3848 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3480..3499) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3690..3711 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3865..3884) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3984..4006 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4044..4062) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4130..4234 /gene="bla" /label=AmpR promoter CDS 4235..5095 /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(4453..4472) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5266..5854 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5755..5774 /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 cgcgtctgca gagggccctg cgtatgagtg 181 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc 241 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg 301 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca 361 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg 421 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc 481 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac 541 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga 601 gtcgtgtcgt gcctgagagc gcagacgcgt ggacggtacc cttcaggtcg acgtcgataa 661 cttcgtatag catacattat acgaagttat ccgaatcgca ataacttcgt ataggatact 721 ttatacgaag ttatttggcg cgccgctagc ttacttgtac agctcgtcca tgccgtacag 781 aaacagatgg tgcctcccct cgctccgctc atactgctcc acaatggtgt agtcttcgtt 841 atggctagta atatccagct ttgtgtcgac gtaatagtag ccaggcagct gcacgggttt 901 cttggccatg tagatggttt tgaattcgac cagataatgc cctccatctt tcagcttcag 961 tgcctggtgg atctctccct tcagcacgcc gtctctgggg tacagccttt ctgttgaagc 1021 ctcccagccc attgtcttct tctgcatgac tgggccatca gggggaaaat tggtgccgcg 1081 cattttcacc ttatagatca gagtgccgtc ctgcagagag ctgtcctggg tgacagtcac 1141 caggcccccg tcttcgaagt tcatcactct ttcccactta aacccctctg ggaagctcag 1201 tttcttgtaa tctggaatgt cagcagggtg tttcacgtag gctttacttc catacataaa 1261 ctgaggagac aggatgtccc atgcgaatgg cagagggccg ccctttgtca ctttcagctt 1321 tgctgtctgg gtgccctcat agggcctccc ttccccctct ccttcaatct caaactcgtg 1381 cccgttcatg ctgccctcca tcctgacttt aaacctcata aactccttaa tcacggccat 1441 gttgttgtcc tcggaggagg cggtgccgga gctgccgctg ccggtgctgc cggtaccatg 1501 ccccaggaac aggtggtggc ggccctcgga gcgctcgtac tgttccacga tggtgtagtc 1561 ctcgttgtgg gaggtgatgt ccagcttggt gtccacgtag tagtagccgg gcagttgcac 1621 gggcttcttg gccatgtaga tggtcttgaa ctccaccagg tagtggccgc cgtccttcag 1681 cttcagggcc tggtggatct cgcccttcag cacgccgtcg cgggggtaca ggcgctcggt 1741 ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggggg ggaagttggt 1801 gccgcgcatc ttcaccttgt agatcagcgt gccgtcctgc agggaggagt cctgggtcac 1861 ggtcaccaga ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa 1921 ggacagcttc ttgtaatcgg ggatgtcggc ggggtgcttc acgtacgcct tggagccgta 1981 catgaactgg ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctt 2041 cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa 2101 ctcgtggccg ttcatggagc cctccatgcg caccttgaag cgcatgaact ctttgatgac 2161 ctcctcgccc ttgctcacca tggtggcgct agcggcgcgc caacctggcc ggccttgata 2221 acttcgtata atgtatgcta tacgaagtta tccgaatcgc aataacttcg tataaagtat 2281 cctatacgaa gttatcgaag atcatcaagc ttatcgataa tcaacctctg gattacaaaa 2341 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 2401 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2461 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2521 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2581 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2641 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 2701 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc 2761 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 2821 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 2881 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatcgatct 2941 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 3001 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 3061 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 3121 gaggattggg aagacaatag caggcatgct ggggacacgt gcggaccgag cggccgcagg 3181 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 3241 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 3301 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 3361 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc 3421 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 3481 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 3541 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 3601 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 3661 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 3721 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg 3781 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 3841 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc 3901 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 3961 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 4021 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 4081 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 4141 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 4201 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 4261 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 4321 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 4381 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 4441 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 4501 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 4561 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 4621 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 4681 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 4741 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 4801 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 4861 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 4921 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 4981 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 5041 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 5101 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 5161 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 5221 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 5281 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 5341 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 5401 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 5461 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 5521 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 5581 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 5641 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 5701 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 5761 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 5821 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 5881 gttcctggcc ttttgctggc cttttgctca catgt //