LOCUS Exported 5740 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pAAV plasmid expressing an NOS-IN133.3xFLAG.mCherry fusion protein under the hSyn promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-NOS-IN133.3xFLAG.mCherry-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5740) AUTHORS Candemir E, Kollert L, Weissflog L, Geis M, Muller A, Post AM, O'Leary A, Harro J, Reif A, Freudenberg F TITLE Interaction of NOS1AP with the NOS-I PDZ domain: Implications for schizophrenia-related alterations in dendritic morphology. JOURNAL Eur Neuropsychopharmacol. 2016 Apr;26(4):741-55. doi: 10.1016/j.euroneuro.2016.01.008. Epub 2016 Jan 28. PUBMED 26861996 REFERENCE 2 (bases 1 to 5740) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5740 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 660..669 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1074..1139 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1146..1856 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1291..1309) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1559..1578 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1867..2455 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1920..1940) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2338..2349) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2487..2963 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2686..2705) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3003..3143 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3003..3132 /label=AAV2 ITR rep_origin 3218..3673 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3305..3324) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3515..3536 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3690..3709) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3809..3831 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3869..3887) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3955..4059 /gene="bla" /label=AmpR promoter CDS 4060..4920 /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(4278..4297) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5091..5679 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5580..5599 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacactagtg 661 ccaccatgga agagcacacg tttggggtcc agcagatcca acccaacgtc atttctgtcc 721 gtctcttcaa acgcaaagtg ggaggtctgg gcttcctggt gaaggaacgg gtcagcaagc 781 ccccggtgat catctcagat ctgatccgag gaggtgccgc ggagcagagc ggccttatcc 841 aagccggcga catcattctc gccgtcaatg atcggcccct ggtagacctc agctatgaca 901 gcgctctgga agttctcagg ggcattgcct ctgagaccca tgtggtcctc attctgagag 961 gccctgaggg ctttactaca cacctggaga ccaccttcac aggggatgga acccccaaga 1021 ccatccgtgt gacccagccc ctggggacac ccaccaaagc tgtcgatgaa ttcgactaca 1081 aagaccatga cggtgattat aaagatcatg atattgatta caaggatgac gatgacaagc 1141 tagctatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct 1201 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg 1261 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc 1321 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga 1381 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg 1441 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc 1501 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc 1561 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg 1621 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg 1681 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca 1741 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt 1801 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagctt 1861 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 1921 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 1981 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 2041 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 2101 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 2161 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 2221 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg 2281 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 2341 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 2401 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga 2461 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc 2521 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt 2581 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat 2641 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa 2701 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga 2761 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct 2821 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact 2881 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa 2941 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc 3001 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 3061 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 3121 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg 3181 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta 3241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg 3301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3421 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt 3481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 3541 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc 3601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 3661 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 3721 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 3781 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 3841 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 3901 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 3961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4021 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 4081 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 4141 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 4201 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 4261 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 4321 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 4381 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 4441 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 4501 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 4561 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 4621 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 4681 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 4741 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 4801 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 4861 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 4921 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 4981 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 5041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 5101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 5161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 5221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 5281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 5341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 5401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 5461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 5521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 5581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 5641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 5701 ttacggttcc tggccttttg ctggcctttt gctcacatgt //