LOCUS Exported 5810 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV TALE synthesis backbone with NG half repeat; fused to nuclear localization sequence linker and SID4X transcriptional repressor effector domain. Synapsin promoter for neuronal expression.. ACCESSION . VERSION . KEYWORDS pAAV_hSyn_TALEBB(NG)-NLS-SID4X_2A_phiLOV2.1_WPRE_bGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5810) AUTHORS Konermann S, Brigham MD, Trevino A, Hsu PD, Heidenreich M, Le Cong, Platt RJ, Scott DA, Church GM, Zhang F TITLE Optical control of mammalian endogenous transcription and epigenetic states. JOURNAL Nature. 2013 Jul 23. doi: 10.1038/nature12466. PUBMED 23877069 REFERENCE 2 (bases 1 to 5810) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5810 /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 181..628 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 651..677 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1370..1390 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1790..1843 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" misc_feature 2201..2789 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2254..2274) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2672..2683) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2831..2848) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2837..3044 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3073..3213 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3073..3202 /label=AAV2 ITR rep_origin 3288..3743 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3375..3394) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3585..3606 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3760..3779) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3879..3901 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3939..3957) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4025..4129 /gene="bla" /label=AmpR promoter CDS 4130..4990 /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(4348..4367) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5161..5749 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5650..5669 /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 gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg 181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tacccatacg 661 atgttccaga ttacgctgta gatttgagaa ctttgggata ttcacagcag cagcaggaaa 721 agatcaagcc caaagtgagg tcgacagtcg cgcagcatca cgaagcgctg gtgggtcatg 781 ggtttacaca tgcccacatc gtagccttgt cgcagcaccc tgcagccctt ggcacggtcg 841 ccgtcaagta ccaggacatg attgcggcgt tgccggaagc cacacatgag gcgatcgtcg 901 gtgtggggaa acagtggagc ggagcccgag cgcttgaggc cctgttgacg gtcgcgggag 961 agctgagagg gcctcccctt cagctggaca cgggccagtt gctgaagatc gcgaagcggg 1021 gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa tgcgctcacg ggagcacccc 1081 tcaacctgac agagacccgt acgggtctcg actcacgcct gagcaggtag tggctattgc 1141 atccaatggc gggggcagac ccgcactgga gtcaatcgtg gcccagcttt cgaggccgga 1201 ccccgcgctg gccgcactca ctaatgatca tcttgtagcg ctggcctgcc tcggcggacg 1261 acccgccttg gatgcggtga agaaggggct cccgcacgcg cctgcattga ttaagcggac 1321 caacagaagg attcccgaga ggacatcaca tcgagtggca gctagcagcc ccaagaagaa 1381 gagaaaggtg gaggccagca tgaacatcca gatgctgctg gaggccgctg actacctgga 1441 acggagagag cgcgaagccg agcacggata tgcttcaatg ctgcccggaa gcggcatgaa 1501 tattcagatg ctgctggagg ctgctgatta cctggaaagg cgcgaacggg aggccgaaca 1561 tggctatgct tccatgctgc ctgggtctgg aatgaatatc caaatgctgc tggaggcagc 1621 cgattacctg gaacggagag aaagagaagc cgagcacgga tacgccagca tgctgccagg 1681 cagcgggatg aacatacaaa tgctgctgga ggctgccgat tacctggaga ggcgcgagag 1741 agaagctgaa catggctatg cctctatgct gcccccatgg ggcagtggag agggcagagg 1801 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaatcgaga agtccttcgt 1861 aataacagat ccaagactgc ctgattaccc catcattttt gccagcgacg gttttctgga 1921 gctcaccgaa tattccaggg aggaaatcat ggggagaaac gccaggttcc tgcaagggcc 1981 tgagacagac caggccaccg ttcagaagat tcgcgatgct atccgcgatc agagggaaac 2041 cactgtgcag ttaataaact acactaagtc aggcaaaaag ttctggaatc tgcttcatct 2101 ccaacctgtc cgggatcgaa agggagggct gcaatacttc atcggtgtgc agctggtggg 2161 aagtgatcat gtgtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa 2221 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 2281 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 2341 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 2401 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 2461 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 2521 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 2581 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat 2641 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 2701 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 2761 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agctagagct 2821 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2881 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2941 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3001 agcaaggggg aggattggga agacaatagc aggcatgctg gggaggtaac cacgtgcgga 3061 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 3121 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 3181 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta 3241 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag 3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 3421 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 3481 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata 3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 3601 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc 3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 3721 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc 3781 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3901 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3961 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 4021 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 4081 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 4141 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 4201 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 4261 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 4321 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 4381 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 4441 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 4501 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 4561 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 4621 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 4681 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 4741 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 4801 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat 4861 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 4921 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 4981 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 5041 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 5101 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 5161 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 5221 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 5281 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 5341 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 5401 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 5461 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 5521 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 5581 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 5641 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 5701 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 5761 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt //