LOCUS Exported 6271 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION high transduction efficiency AAV-mediated synapsin promoter-dependent expression of AICD (last 50 amino acids of APP) and IRES-mediated co-expression of hrGFP . ACCESSION . VERSION . KEYWORDS pAAV-syn-AICD-IRES-hrGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6271) AUTHORS Pousinha PA, Mouska X, Raymond EF, Gwizdek C, Dhib G, Poupon G, Zaragosi LE, Giudici C, Bethus I, Pacary E, Willem M, Marie H TITLE Physiological and pathophysiological control of synaptic GluN2B-NMDA receptors by the C-terminal domain of amyloid precursor protein. JOURNAL Elife. 2017 Jul 6;6. doi: 10.7554/eLife.25659. PUBMED 28682239 REFERENCE 2 (bases 1 to 6271) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6271 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 22..151 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 362..809 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 822..831 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 990..1061 /codon_start=1 /product="three tandem FLAG(R) epitope tags" /label=3xFLAG /translation="DYKDDDDKDYKDDDDKDYKDDDDK" misc_feature 1097..1680 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1264..1281) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1491..1510 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1681..2400 /codon_start=1 /product="humanized Renilla green fluorescent protein" /label=hrGFP /note="mammalian codon-optimized" /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI AQLTSLGKPLGSLHEWV" misc_feature 2425..3013 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2478..2498) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2896..2907) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3033..3509 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3232..3251) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3555..3695 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3555..3684 /label=AAV2 ITR rep_origin 3770..4225 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3857..3876) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4067..4088 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4242..4261) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4361..4383 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4421..4439) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4507..4611 /gene="bla" /label=AmpR promoter CDS 4612..5472 /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(4830..4849) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5643..6231 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6132..6151 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg 61 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag 121 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgttgcaa agatggataa 181 agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg 241 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 301 tggggggagg ggtcggcagc aaatggttaa ttaatctaga ctgcagaggg ccctgcgtat 361 gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg 421 accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag 481 agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc 541 ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg 601 cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc 661 gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc 721 gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg 781 gaggagtcgt gtcgtgcctg agagcgcagt cgagaggatc cgccaccatg gtgatgctga 841 agaagaaaca gtacacatcc attcatcatg gtgtggtgga ggttgacgcc gctgtcaccc 901 cagaggagcg ccacctgtcc aagatgcagc agaacggcta cgaaaatcca acctacaagt 961 tctttgagca gatgcagaac tgactcgagg actacaagga tgacgatgac aaggattaca 1021 aagacgacga tgataaggac tataaggatg atgacgacaa ataatagcaa ttcctcgacg 1081 actgcatagg gttacccccc tctccctccc ccccccctaa cgttactggc cgaagccgct 1141 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg 1201 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt 1261 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg 1321 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac 1381 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg 1441 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct 1501 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg 1561 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg 1621 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat ggccacaacc 1681 atggtgagca agcagatcct gaagaacacc ggcctgcagg agatcatgag cttcaaggtg 1741 aacctggagg gcgtggtgaa caaccacgtg ttcaccatgg agggctgcgg caagggcaac 1801 atcctgttcg gcaaccagct ggtgcagatc cgcgtgacca agggcgcccc cctgcccttc 1861 gccttcgaca tcctgagccc cgccttccag tacggcaacc gcaccttcac caagtacccc 1921 gaggacatca gcgacttctt catccagagc ttccccgccg gcttcgtgta cgagcgcacc 1981 ctgcgctacg aggacggcgg cctggtggag atccgcagcg acatcaacct gatcgaggag 2041 atgttcgtgt accgcgtgga gtacaagggc cgcaacttcc ccaacgacgg ccccgtgatg 2101 aagaagacca tcaccggcct gcagcccagc ttcgaggtgg tgtacatgaa cgacggcgtg 2161 ctggtgggcc aggtgatcct ggtgtaccgc ctgaacagcg gcaagttcta cagctgccac 2221 atgcgcaccc tgatgaagag caagggcgtg gtgaaggact tccccgagta ccacttcatc 2281 cagcaccgcc tggagaagac ctacgtggag gacggcggct tcgtggagca gcacgagacc 2341 gccatcgccc agctgaccag cctgggcaag cccctgggca gcctgcacga gtgggtgtaa 2401 gaattccgta caaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg 2461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 2521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 2581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 2641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 2701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 2761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 2821 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc 2881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 2941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 3001 gccgcctccc cgcatcgata ccggctcgag acgggtggca tccctgtgac ccctccccag 3061 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat 3121 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg 3181 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg 3241 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc 3301 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc 3361 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct 3421 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg 3481 cgtgaaccac tgctcccttc cctgtccttc tcgagctgat tttgtaggta accacgtgcg 3541 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 3601 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 3661 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 3721 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3841 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 3961 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 4021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 4081 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg 4141 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 4201 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 4261 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 4321 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 4381 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 4441 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 4501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 4561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 4621 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 4681 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 4741 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 4801 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 4861 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 4921 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 4981 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 5041 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 5101 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 5161 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 5221 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 5281 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 5341 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 5401 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 5461 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 5521 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5581 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5641 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5701 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5761 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5821 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5881 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5941 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 6001 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 6061 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 6121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 6181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 6241 aacgcggcct ttttacggtt cctggccttt t //