LOCUS Exported 6335 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Targeting expression to neuronal and glial cell types in mice, non-human primates, and human retina. ACCESSION . VERSION . KEYWORDS 179_pAAV-ProD10-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6335) 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 6335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6335 /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 694..725 /label=minP /note="minimal TATA-box promoter with low basal activity" regulatory 779..788 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 785..1708 /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 1715..2431 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1760..1781) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2021..2040) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2368..2389 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2462..3050 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2515..2535) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2933..2944) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3082..3558 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3281..3300) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3598..3738 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3598..3727 /label=AAV2 ITR rep_origin 3813..4268 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3900..3919) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4110..4131 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4285..4304) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4404..4426 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4464..4482) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4550..4654 /gene="bla" /label=AmpR promoter CDS 4655..5515 /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(4873..4892) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5686..6274 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6175..6194 /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 gcgtatcctg gggtttaaac aggactaggt ccactgggag 181 aagaggaagt caaacaggac tagtcatcga taggctagcc agcttaatta atcagatagg 241 aagactagtc agcatcagga aagcttacta cagtatgtat ggtgtgctag tgagctagag 301 ctagagcaaa gccggattac ctccaaacat gctggggaag aattgactac acttctacct 361 aaacagtatg ccagaagaga aacagaagtg tggtgccttc ttagcgtgac tcagagtacc 421 tatgaaagct actagcccgt gttacaagtg actagaaaca ctcaggcatc ggtgctgaaa 481 tcggccaagg agacctgtgt caccatttct gtttagaatc cagtaaaacc agcttagttg 541 acattacagc tcaggcttgg gttgatttct aaagttcaga gccaatcaaa agtcaccaga 601 aaagcccaga gcctgggtgt gttgtgttgc tcagtggtcg agctcctgct gggaggatgg 661 ggagggatcc tcacatggtc ctgctggagt tagtagaggg tatataatgg aagctcgact 721 tccagctatc acatccactg tgttgttgtg aactggaatc cactataggc caagatccgc 781 caccatggat tatggaggcg ccctgagtgc cgttgggcgc gagctgctat ttgtaacgaa 841 cccagtagtc gtcaatggct ctgtacttgt gcctgaggac cagtgttact gcgcgggctg 901 gattgagtcg cgtggcacaa acggtgccca aacggcgtcg aacgtgctgc aatggcttgc 961 tgctggcttc tccatcctac tgcttatgtt ttacgcctac caaacatgga agtcaacctg 1021 cggctgggag gagatctatg tgtgcgctat cgagatggtc aaggtgattc tcgagttctt 1081 cttcgagttt aagaacccgt ccatgctgta tctagccaca ggccaccgcg tccagtggtt 1141 gcgttacgcc gagtggcttc tcacctgccc ggtcatttgc attcacctgt caaacctgac 1201 gggcttgtcc aacgactaca gcaggcgcac catgggtctg cttgtgtctg atattggcac 1261 aattgtgtgg ggcgccactt ccgccatggc caccggatac gtcaaggtca tcttcttctg 1321 cctgggtctg tgttatggtg ctaacacgtt ctttcacgct gccaaggcct acatcgaggg 1381 ttaccacacc gtgccgaagg gccggtgtcg ccaggtggtg actggcatgg cttggctctt 1441 cttcgtatca tggggtatgt tccccatcct gttcatcctc ggccccgagg gcttcggcgt 1501 cctgagcgtg tacggctcca ccgtcggcca caccatcatt gacctgatgt cgaagaactg 1561 ctggggtctg ctcggccact acctgcgcgt gctgatccac gagcatatcc tcatccacgg 1621 cgacattcgc aagaccacca aattgaacat tggtggcact gagattgagg tcgagacgct 1681 ggtggaggac gaggccgagg ctggcgcggt acccatggtg agcaagggcg aggagctgtt 1741 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag 1801 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg 1861 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt 1921 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat 1981 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac 2041 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat 2101 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca 2161 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg 2221 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat 2281 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag 2341 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 2401 gatcactctc ggcatggacg agctgtacaa gtgataagaa ttcgatatca agcttatcga 2461 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2521 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2581 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2641 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2701 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2761 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2821 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 2881 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 2941 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3001 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 3061 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg 3121 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc 3181 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc 3241 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg 3301 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc 3361 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt 3421 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa 3481 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact 3541 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg 3601 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 3661 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 3721 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta 3781 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc 3841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 3901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 3961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 4021 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 4081 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 4141 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg 4201 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 4261 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc 4321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 4381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 4441 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 4501 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 4561 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 4621 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 4681 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 4741 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 4801 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 4861 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 4921 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 4981 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 5041 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 5101 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 5161 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 5221 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 5281 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 5341 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 5401 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 5461 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 5521 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 5581 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 5641 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 5701 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 5761 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 5821 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 5881 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 5941 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 6001 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 6061 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 6121 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 6181 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 6241 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 6301 gttcctggcc ttttgctggc cttttgctca catgt //