LOCUS Exported 6926 bp ds-DNA circular SYN 29-OCT-2021 DEFINITION This AAV plasmid vector expresses human 2N4R microtubule-associated protein tau with P301L mutation.. ACCESSION . VERSION . KEYWORDS pAAV-FLEX-P301L Tau SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6926) AUTHORS You Y, Botros MB, Enoo AAV, Bockmiller A, Herron S, Delpech JC, Ikezu T TITLE Cre-inducible Adeno Associated Virus-mediated Expression of P301L Mutant Tau Causes Motor Deficits and Neuronal Degeneration in the Substantia Nigra. JOURNAL Neuroscience. 2019 Dec 1;422:65-74. doi: 10.1016/j.neuroscience.2019.10.001. Epub 2019 Nov 2. PUBMED 31689387 REFERENCE 2 (bases 1 to 6926) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 29, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6926 /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 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1450..1483 /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 1534..1567 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 2898..2907 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2912..2945) /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(2996..3029) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3054..3642 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3107..3127) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3525..3536) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3674..4149 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3872..3891) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4189..4329 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4189..4318 /label=AAV2 ITR rep_origin 4404..4859 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4491..4510) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4701..4722 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4876..4895) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4995..5017 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5055..5073) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5141..5245 /gene="bla" /label=AmpR promoter CDS 5246..6106 /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(5464..5483) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6277..6865 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6766..6785 /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 cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc 1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga 1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg 1561 aagttatggc gcgcctcaca aaccctgctt ggccagggag gcagacacct cgtcagctag 1621 cgtggcgagc tggggcgagt ctaccatgtc gatgctgccg gtggaggaga cattgctgag 1681 atgccgtgga gacgtgtccc cagacaccac tggcgacttg tacacgatct ccgccccgtg 1741 gtctgtcttg gctttggcgt tctcgcggaa ggtcagcttg tgggtttcaa tctttttatt 1801 tcctccgcca gggacgtggg tgatattgtc cagggaccca atcttcgact ggactctgtc 1861 cttgaagtca agcttctcag attttacttc cacctggcca cctcctggtt tatgatggat 1921 gttgcctaat gagccacact tggaggtcac cttgctcagg tcaactggtt tgtagactat 1981 ttgcacactg ccgcctccca ggacgtgttt gatattatcc tttgagccac acttggactg 2041 gacgttgcta agatccagct tcttattaat tatctgcacc ttcccgcctc ccggctggtg 2101 cttcaggttc tcagtggagc cgatcttgga cttgacattc ttcaggtctg gcatgggcac 2161 gggggctgtc tgcaggcggc tcttggcgga agacggcgac ttgggtggag tacggaccac 2221 tgccaccttc ttgggctccc gggtgggtgg ggttggaagg gacggggtgc gggagcggct 2281 gccgggagtg cctggggagc cggggctgct gtagccgctg cgatcccctg attttggagg 2341 ttcaccagag ctgggtggtg tctttggagc gggcggggtt tttgctggaa tcctggtggc 2401 gttggcctgg cccttctggc ctggaggggc tgctccccgc ggtgtggcga tcttcgtttt 2461 accatcagcc cccttggctt ttttgtcatc gcttccagtc ccgtctttgc ttttactgac 2521 catgcgagct tgggtcacgt gaccagcagc ttcgtcttcc aggctggggg tgtctccaat 2581 gcctgcttct tcagctgtgg ttccttctgg gatctccgtg tggggctgcg cggcagcctg 2641 cttgccggga gctccctcat ccactaaggg tgctgtcaca tcttccgctg ttggagtgct 2701 cttagcatca gaggtttcag agcccggttc ctcagatccg tcctcagtgg gggtctgcag 2761 gggagattct ttcaggccag cgtccgtgtc accctcttgg tcttggtgca tggtgtagcc 2821 cccctgatct ttcctgtccc ccaacccgta cgtcccagcg tgatcttcca tcacttcgaa 2881 ctcctggcgg ggctcagcca tggtggctag cataacttcg tataaagtat cctatacgaa 2941 gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc aaagaataac 3001 ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc gataatcaac 3061 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 3121 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 3181 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 3241 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 3301 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 3361 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 3421 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg 3481 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 3541 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 3601 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc 3661 tcgagagatc tacgggtggc atccctgtac ccctccccag tgcctctcct ggccctggaa 3721 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct 3781 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa 3841 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg 3901 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag 3961 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt 4021 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg 4081 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc 4141 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag 4201 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4261 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct 4321 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 4381 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 4441 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 4501 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 4561 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt 4621 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 4681 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 4741 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 4801 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt 4861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 4921 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 4981 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 5041 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 5101 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 5161 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 5221 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 5281 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 5341 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 5401 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 5461 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 5521 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 5581 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 5641 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 5701 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 5761 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 5821 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 5881 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 5941 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 6001 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 6061 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 6121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 6181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 6241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 6301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 6361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 6421 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 6481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 6541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 6601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 6661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 6721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 6781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 6841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 6901 cttttgctgg ccttttgctc acatgt //