LOCUS Exported 6302 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV-mediated expression of last 50 amino acids of APLP1 protein (APLP1ICD) and IRES-mediated co-expression of hrGFP to recognize labelled cells. ACCESSION . VERSION . KEYWORDS pAAV-APLP1ICD-IRES-hrGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6302) 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 6302) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6302 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 246..549 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 550..753 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 704..724 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 750..774 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1351..1369 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 1622..1693 /codon_start=1 /product="three tandem FLAG(R) epitope tags" /label=3xFLAG /translation="DYKDDDDKDYKDDDDKDYKDDDDK" misc_feature 1729..2312 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1896..1913) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2123..2142 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2313..3032 /codon_start=1 /product="humanized Renilla green fluorescent protein" /label=hrGFP /note="mammalian codon-optimized" /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI AQLTSLGKPLGSLHEWV" primer_bind complement(3062..3081) /label=T7 /note="T7 promoter, forward primer" promoter complement(3063..3081) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 3089..3565 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3288..3307) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3605..3745 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3605..3734 /label=AAV2 ITR rep_origin 3820..4275 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3907..3926) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4117..4138 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4292..4311) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4411..4433 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4471..4489) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4557..4661 /gene="bla" /label=AmpR promoter CDS 4662..5522 /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(4880..4899) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5693..6281 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6182..6201 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct 61 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag 121 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac 181 gcgtctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 241 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 301 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 361 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 421 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 481 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 541 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 601 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 661 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 721 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 781 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc 841 ggattcgaat cccggccggg aacggtgcat tggaacgcgg attccccgtg ccaagagtga 901 cgtaagtacc gcctatagag tctataggcc cacaaaaaat gctttcttct tttaatatac 961 ttttttgttt atcttatttc taatactttc cctaatctct ttctttcagg gcaataatga 1021 tacaatgtat catgcctctt tgcaccattc taaagaataa cagtgataat ttctgggtta 1081 aggcaatagc aatatttctg catataaata tttctgcata taaattgtaa ctgatgtaag 1141 aggtttcata ttgctaatag cagctacaat ccagctacca ttctgctttt attttatggt 1201 tgggataagg ctggattatt ctgagtccaa gctaggccct tttgctaatc atgttcatac 1261 ctcttatctt cctcccacag ctcctgggca acgtgctggt ctgtgtgctg gcccatcact 1321 ttggcaaaga attgggattc gaacatcgat aattaaccct cactaaaggg aacaaaagct 1381 ggagctccac cgcggtggcg gccgctctag cccgggcgga tccgaattcg ccaccatgct 1441 gctcctgcgc aggaagaagc cctacggggc tatcagccat ggcgtggtgg aggtggaccc 1501 catgctgacc ctggaggagc agcagctccg cgaactgcag cggcacggct atgagaaccc 1561 cacttaccgc ttcctggagg aacgaccctg actcttcctg gaggaacgac cctgactcga 1621 ggactacaag gatgacgatg acaaggatta caaagacgac gatgataagg actataagga 1681 tgatgacgac aaataatagc aattcctcga cgactgcata gggttacccc cctctccctc 1741 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta 1801 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc 1861 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct 1921 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt 1981 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa 2041 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg 2101 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga 2161 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac 2221 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc 2281 ctttgaaaaa cacgatgata atggccacaa ccatggtgag caagcagatc ctgaagaaca 2341 ccggcctgca ggagatcatg agcttcaagg tgaacctgga gggcgtggtg aacaaccacg 2401 tgttcaccat ggagggctgc ggcaagggca acatcctgtt cggcaaccag ctggtgcaga 2461 tccgcgtgac caagggcgcc cccctgccct tcgccttcga catcctgagc cccgccttcc 2521 agtacggcaa ccgcaccttc accaagtacc ccgaggacat cagcgacttc ttcatccaga 2581 gcttccccgc cggcttcgtg tacgagcgca ccctgcgcta cgaggacggc ggcctggtgg 2641 agatccgcag cgacatcaac ctgatcgagg agatgttcgt gtaccgcgtg gagtacaagg 2701 gccgcaactt ccccaacgac ggccccgtga tgaagaagac catcaccggc ctgcagccca 2761 gcttcgaggt ggtgtacatg aacgacggcg tgctggtggg ccaggtgatc ctggtgtacc 2821 gcctgaacag cggcaagttc tacagctgcc acatgcgcac cctgatgaag agcaagggcg 2881 tggtgaagga cttccccgag taccacttca tccagcaccg cctggagaag acctacgtgg 2941 aggacggcgg cttcgtggag cagcacgaga ccgccatcgc ccagctgacc agcctgggca 3001 agcccctggg cagcctgcac gagtgggtgt aatagggtac caggtaagtg tacccaattc 3061 gccctatagt gagtcgtatt agatctacgg gtggcatccc tgtgacccct ccccagtgcc 3121 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag 3181 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt 3241 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc 3301 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc 3361 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag 3421 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa 3481 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg 3541 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg 3601 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact 3661 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc 3721 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg 3781 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat 3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag 3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 4021 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt 4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 4141 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg 4201 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 4261 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 4321 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 4381 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 4441 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta 4501 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg 4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 5521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 5701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 5761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 5821 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 5881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 5941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 6001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 6061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 6121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 6181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 6241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 6301 tt //