LOCUS Exported 5749 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Global labeling of excitatory synapses (green fluorescence). ACCESSION . VERSION . KEYWORDS AAV-Syn-PSD95.FingR-eGFP-CCR5TC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5749) AUTHORS Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man HY, Han X TITLE A Viral Toolbox of Genetically Encoded Fluorescent Synaptic Tags. JOURNAL iScience. 2020 Jun 30;23(7):101330. doi: 10.1016/j.isci.2020.101330. PUBMED 32674057 REFERENCE 2 (bases 1 to 5749) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5749 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR protein_bind 196..207 /label=ZF binding site /bound_moiety="CCR5TC zinc-finger domain" /note="target sequence for the CCR5TC zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 2008; Gross et al., 2013)" promoter 235..682 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 986..1009 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1034..1744 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" CDS 1034..1744 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" primer_bind complement(1076..1097) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1337..1356) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1684..1705 /label=EGFP-C /note="EGFP, forward primer" CDS 1790..2131 /codon_start=1 /product="zinc-finger domain from the CCR5-224 (+) zinc-finger nuclease that recognizes the left half-site target sequence in the human CCR5 gene (Perez et al., 2008; Gross et al., 2013)" /label=CCR5TC /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH TKIHLRGSQL" CDS 2180..2305 /codon_start=1 /product="Kruppel-associated box (KRAB) transcriptional repression domain from the rat zinc finger protein Kid-1 (Witzgall et al., 1994)" /label=KRAB-A /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM" misc_feature 2336..2924 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2389..2409) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2807..2818) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2927..2943) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2928..2944) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 2973..3094 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2984..3003) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3038..3057 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3302..3319) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3473..4061) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3553..3572) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4232..5092) /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 4855..4874 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5093..5197) /gene="bla" /label=AmpR promoter rep_origin complement(5223..5678) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5360..5381) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5572..5591 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 181 aggaagatca cgcgtgtcat cctcatcaga tctctgcaga gggccctgcg tatgagtgca 241 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 301 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 361 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 421 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 481 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 541 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 601 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 661 cgtgtcgtgc ctgagagcgc agtcgaattc aagctgctag caaggatccg ccaccatgct 721 cgaagtcaag gaagcatcac caaccagcat ccagatcagc tgggtgctcc acttgcgcca 781 cgttcgctac taccgcatca cctacggtga aactggtggc aatagccctg tccaggaatt 841 caccgtgcct ggcagcaagt ccactgctac catcagcggc ctgaaacctg gtgtcgacta 901 taccatcacg gtgtacgccg tcacgatctt cagcgcctac cgctccgcct ggccgccgat 961 ctccatcaac taccgcaccg gaaccgatta caaggatgac gacgataagg gtagcggctc 1021 cagtagatct ggggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt 1081 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 1141 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 1201 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga 1261 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 1321 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 1381 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 1441 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa 1501 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt 1561 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 1621 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 1681 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 1741 gtactctagg ggagctggcg ctggagcggg tgcaggggct ggctctagat tccagtgccg 1801 gatctgcatg cggaacttca gcgaccggtc caacctgagc aggcacatca gaacccacac 1861 cggagaaaag cccttcgcct gcgacatttg cggccggaag ttcgccatca gcagcaacct 1921 gaacagccac accaagatcc acactggcag ccagaaacct ttccagtgca gaatttgtat 1981 gagaaacttt agcagaagcg acaacctggc cagacacatc cggacacata ctggtgaaaa 2041 accttttgcc tgtgatatct gtggcagaaa gtttgccacc tccggcaatc tgacccggca 2101 cacaaagatt cacctgcggg gcagccagct atcgattgtc gacgctcctg aacaacgtga 2161 aggtgcttct caagtttctg tttctgttac ttttgaagat gttgctgttc tttttactcg 2221 tgatgaatgg aaaaaacttg atctttctca acgttctctt tatcgtgaag ttatgcttga 2281 aaattattct aatcttgctt ctatggctta agaattcgat atcaagctta tcgataatca 2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 2461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 2521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 2581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 2641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 2701 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg 2761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 2821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 2881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc 2941 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac 3001 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 3061 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 3121 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 3181 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt 3241 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 3301 sctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 3361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 3421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 3481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 3541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 3601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 3661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 3721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 3781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 3841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 3901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 3961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 4021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 4081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 4141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 4201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 4261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 4321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 4381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 4441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 4501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 4561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 4621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 4681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 4741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 4801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 4861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 4921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 4981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 5041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 5101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 5161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 5221 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 5281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 5341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 5401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 5461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 5521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 5581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 5641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 5701 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccag //