LOCUS Exported 7707 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Targeting expression to neuronal and glial cell types in mice, non-human primates, and human retina. ACCESSION . VERSION . KEYWORDS 78_pAAV-ProA27-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7707) AUTHORS Juttner J, Szabo A, Gross-Scherf B, Morikawa RK, Rompani SB, Hantz P, Szikra T, Esposti F, Cowan CS, Bharioke A, Patino-Alvarez CP, Keles O, Kusnyerik A, Azoulay T, Hartl D, Krebs AR, Schubeler D, Hajdu RI, Lukats A, Nemeth J, Nagy ZZ, Wu KC, Wu RH, Xiang L, Fang XL, Jin ZB, Goldblum D, Hasler PW, Scholl HPN, Krol J, Roska B TITLE Targeting neuronal and glial cell types with synthetic promoter AAVs in mice, non-human primates and humans. JOURNAL Nat Neurosci. 2019 Jul 8. pii: 10.1038/s41593-019-0431-2. doi: 10.1038/s41593-019-0431-2. PUBMED 31285614 REFERENCE 2 (bases 1 to 7707) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7707 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 2151..2160 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2157..3080 /codon_start=1 /product="N-terminal fragment of channelrhodopsin-2 from the alga Chlamydomonas reinhardtii (Nagel et al., 2003)" /label=ChR2 /note="blue light-activated cation channel" /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN PSMLYLATGHRVQWLRYAEWLLTCPVICIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGA" CDS 3087..3803 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3132..3153) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3393..3412) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3740..3761 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3834..4422 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3887..3907) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4305..4316) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4454..4930 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4653..4672) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4970..5110 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4970..5099 /label=AAV2 ITR rep_origin 5185..5640 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5272..5291) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5482..5503 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5657..5676) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5776..5798 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5836..5854) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5922..6026 /gene="bla" /label=AmpR promoter CDS 6027..6887 /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(6245..6264) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7058..7646 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7547..7566 /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 gcgtggtcag taaagtgaat gagatgaggt catgttccat 181 gtgggataaa catggttagt agactgcacc aggtgatggg ggaaaggaga gaaaaagatg 241 tgagagggga ggggaccagg agaagggacc aagagcgaag agaatcaaga tagccaagag 301 agcaaatggc caaaaatggc agggttacat aggaaagaga agcttgcaga agagtagcca 361 gggccccagg gaggagatgt ttagttcagg ggagaggtga gaaaaactga gaagagctac 421 aggtactgat ggaagataga agccagaatg aattttggtg tgctaatagg taccacagtt 481 agccagttgc ttctttggaa cccaacataa aggacttttt ttcctcctcc tcctcctcct 541 cctcctcttc ctcttcctct tcctcctcct cctcttcctt ctctctctct ctctctctct 601 ctctctctct ctctcataca taggtgtagc tagaaactgg agcactgtga ccttattcaa 661 ggggccacaa aggtgcctct acacccccat gcacacacct tacaaaaaag gatctctagc 721 atttttgcag tctttcagtg gaattcaaaa aggaatcatt tgattggtaa tgagaatgtt 781 tttcctctcg gcttttcaca gctcaggaag gagtaattta tcaagtactc tgcttggaga 841 agtagagcat agcctggaat ggaggaacat ggatccctgt gggcgcttgc aatcttctgc 901 tttcattttg tcagtcatgc aagcacagtt ttaagtactc accaggggag aaaaagctct 961 cttcccagct tgtgcttgcc agtaagacac tctcctttca gagtcaaggc tgcagctggc 1021 cttgggagct tcttcctagg tctctagcaa gtggagccag agctgctaca gaggaggtaa 1081 gggccatggg cagcatccca ggaaggtggt tggcaagagg gtgggagacc tctctgcacg 1141 tcctcctggg acctgcatgc tctccccatg cctcaccctc accctagttg tgacaccaga 1201 ccaagaaagg cgatgtggtt ccccagccac atccaaagcc aagcctttgg ccagaggaag 1261 gaaggctgtc ccagtcatgg ttttgctttt gtttttcccc tcagcttgct ttttgaacct 1321 ctatctgaga ggacagtgcc ctgctgctcc atagcttcaa tcctggctgc agggatggag 1381 gttgggaagg aagctggtga gatcctagat cctcaaaaat gataactgga tgtagccctg 1441 gtagctaaac atccacagat acacagctgt gaatggcctg tacttggaca aactggagtt 1501 ctgagtttag gatccagatg gattaggagt gagtcccaag caggacttgc ttacccactt 1561 cagggaccag cttccttcca aatagacaca ggagagtgtc tctcttccct gtcacccata 1621 gcctgactag ggagagtgag tggcaaaccc aaatttgcat atttcattcc agctgagact 1681 aaagccttga cctcaggcat ggaggactgt aaccactgag aactaatatg ataacattgt 1741 tatcttacga gaattctcct gtatgcagag taggtccata tggttttctg gattcattgc 1801 acagatgaat tcgaacctga catcccacag ggagagagat gtgtgagctc cagacagaag 1861 gctgtgtagt gtgggcactg tgccaggacc accagcataa tttctcctaa gacacacctg 1921 tctagaaagt tagagccata agacaacctt ggccctgggc catgtcactg ctcatatctg 1981 gccccagctt tgtgtgccta gaggagtatt cttgcctctt tggaaaaatt ggagggagcc 2041 ggattaacct actgcaggtg ctaggggtgg gtggcaaagc tgtgataggt gttggtagtg 2101 ttgtagctgg caaagctgat ctcctgtctg ttgcagcctt caccagatcc gccaccatgg 2161 attatggagg cgccctgagt gccgttgggc gcgagctgct atttgtaacg aacccagtag 2221 tcgtcaatgg ctctgtactt gtgcctgagg accagtgtta ctgcgcgggc tggattgagt 2281 cgcgtggcac aaacggtgcc caaacggcgt cgaacgtgct gcaatggctt gctgctggct 2341 tctccatcct actgcttatg ttttacgcct accaaacatg gaagtcaacc tgcggctggg 2401 aggagatcta tgtgtgcgct atcgagatgg tcaaggtgat tctcgagttc ttcttcgagt 2461 ttaagaaccc gtccatgctg tatctagcca caggccaccg cgtccagtgg ttgcgttacg 2521 ccgagtggct tctcacctgc ccggtcattt gcattcacct gtcaaacctg acgggcttgt 2581 ccaacgacta cagcaggcgc accatgggtc tgcttgtgtc tgatattggc acaattgtgt 2641 ggggcgccac ttccgccatg gccaccggat acgtcaaggt catcttcttc tgcctgggtc 2701 tgtgttatgg tgctaacacg ttctttcacg ctgccaaggc ctacatcgag ggttaccaca 2761 ccgtgccgaa gggccggtgt cgccaggtgg tgactggcat ggcttggctc ttcttcgtat 2821 catggggtat gttccccatc ctgttcatcc tcggccccga gggcttcggc gtcctgagcg 2881 tgtacggctc caccgtcggc cacaccatca ttgacctgat gtcgaagaac tgctggggtc 2941 tgctcggcca ctacctgcgc gtgctgatcc acgagcatat cctcatccac ggcgacattc 3001 gcaagaccac caaattgaac attggtggca ctgagattga ggtcgagacg ctggtggagg 3061 acgaggccga ggctggcgcg gtacccatgg tgagcaaggg cgaggagctg ttcaccgggg 3121 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 3181 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 3241 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 3301 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 3361 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 3421 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3481 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 3541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 3601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 3661 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 3721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 3781 tcggcatgga cgagctgtac aagtgataag aattcgatat caagcttatc gataatcaac 3841 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 3901 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 3961 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 4021 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 4081 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 4141 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 4201 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg 4261 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 4321 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 4381 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc 4441 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga 4501 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc 4561 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca 4621 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt 4681 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca 4741 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt 4801 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt 4861 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt 4921 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta 4981 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca 5041 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc 5101 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 5161 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg 5221 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg 5281 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 5341 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 5401 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 5461 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta 5521 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa 5581 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt 5641 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc 5701 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 5761 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 5821 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 5881 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 5941 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 6001 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 6061 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 6121 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 6181 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 6241 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 6301 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 6361 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 6421 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 6481 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 6541 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 6601 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 6661 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 6721 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 6781 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 6841 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 6901 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 6961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 7021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 7081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 7141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 7201 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 7261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 7321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 7381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 7441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 7501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 7561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 7621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 7681 ccttttgctg gccttttgct cacatgt //