LOCUS Exported 6523 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 214_pAAV-ProD22-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6523) 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 6523) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6523 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 752..783 /label=minP /note="minimal TATA-box promoter with low basal activity" regulatory 837..846 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 843..1766 /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 1773..2489 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1818..1839) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2079..2098) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2426..2447 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2520..3108 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2573..2593) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2991..3002) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3140..3616 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3339..3358) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3656..3796 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3656..3785 /label=AAV2 ITR rep_origin 3871..4326 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3958..3977) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4168..4189 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4343..4362) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4462..4484 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4522..4540) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4608..4712 /gene="bla" /label=AmpR promoter CDS 4713..5573 /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(4931..4950) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5744..6332 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6233..6252 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 6394..6523 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 gcggccgcac gcgtatcctg gggtttaaac aggactaggt ccactgggag aagaggaagt 61 caaacaggac tagtcatcga taggctagcc agcttaatta atcagatatt ccggtcacac 121 ggccaagatt attccacctg cgctttgagc aatagggaga gggctctggt gcctcttcct 181 ggaatttgat taattcgctt gagtcagtca cagaatttga ggaagcattg atatttgaag 241 atgtgttctt ctaaaggata caaatgaata tatgcatagt gagagtttag gagatagggg 301 ttcattgtaa gccactgtag tcgtgggtga ctcacatcaa accacccctt cgggaacacg 361 atgccgacac tgaaactaca taggggagag caaataaaac tgtcttctct agtttgggga 421 aattggagcc tgccttaggg aaacagtgac taatttacag ctagtgtttg gaatgagatc 481 atctgtcaaa ataaatgtat ctttacttcc ttcttgagag gaagcaagct gttttatgat 541 gttccttgga atccttaaaa atacagagca acatttacat tattaatgat acgctttatt 601 gctgcaggct aactaggtcc aaaattgtcc ttccatagat ggagctggag agttacacag 661 aagtttgcat atcgagctct taggtctgca tgtacagcta atgtacttgt ggaccctgtc 721 acatatcctc acatggtcct gctggagtta gtagagggta tataatggaa gctcgacttc 781 cagctatcac atccactgtg ttgttgtgaa ctggaatcca ctataggcca ggatccgcca 841 ccatggatta tggaggcgcc ctgagtgccg ttgggcgcga gctgctattt gtaacgaacc 901 cagtagtcgt caatggctct gtacttgtgc ctgaggacca gtgttactgc gcgggctgga 961 ttgagtcgcg tggcacaaac ggtgcccaaa cggcgtcgaa cgtgctgcaa tggcttgctg 1021 ctggcttctc catcctactg cttatgtttt acgcctacca aacatggaag tcaacctgcg 1081 gctgggagga gatctatgtg tgcgctatcg agatggtcaa ggtgattctc gagttcttct 1141 tcgagtttaa gaacccgtcc atgctgtatc tagccacagg ccaccgcgtc cagtggttgc 1201 gttacgccga gtggcttctc acctgcccgg tcatttgcat tcacctgtca aacctgacgg 1261 gcttgtccaa cgactacagc aggcgcacca tgggtctgct tgtgtctgat attggcacaa 1321 ttgtgtgggg cgccacttcc gccatggcca ccggatacgt caaggtcatc ttcttctgcc 1381 tgggtctgtg ttatggtgct aacacgttct ttcacgctgc caaggcctac atcgagggtt 1441 accacaccgt gccgaagggc cggtgtcgcc aggtggtgac tggcatggct tggctcttct 1501 tcgtatcatg gggtatgttc cccatcctgt tcatcctcgg ccccgagggc ttcggcgtcc 1561 tgagcgtgta cggctccacc gtcggccaca ccatcattga cctgatgtcg aagaactgct 1621 ggggtctgct cggccactac ctgcgcgtgc tgatccacga gcatatcctc atccacggcg 1681 acattcgcaa gaccaccaaa ttgaacattg gtggcactga gattgaggtc gagacgctgg 1741 tggaggacga ggccgaggct ggcgcggtac ccatggtgag caagggcgag gagctgttca 1801 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 1861 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 1921 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 1981 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 2041 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 2101 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 2161 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 2221 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 2281 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 2341 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca 2401 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 2461 tcactctcgg catggacgag ctgtacaagt gataagaatt cgatatcaag cttatcgata 2521 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 2581 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 2641 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 2701 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 2761 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 2821 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 2881 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 2941 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 3001 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 3061 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag 3121 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc 3181 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat 3241 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa 3301 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag 3361 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct 3421 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt 3481 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc 3541 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc 3601 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa 3661 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg 3721 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg 3781 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 3841 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg 3901 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc 3961 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 4021 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 4081 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 4141 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 4201 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt 4261 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta 4321 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 4381 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 4441 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 4501 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 4561 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 4621 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 4681 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 4741 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 4801 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 4861 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 4921 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 4981 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 5041 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 5101 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 5161 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 5221 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 5281 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 5341 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 5401 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 5461 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 5521 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 5581 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 5641 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 5701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 5761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 5821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 5881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 5941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 6001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 6061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 6121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 6181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 6241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 6301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 6361 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc 6421 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga 6481 gcgcgcagag agggagtggc caactccatc actaggggtt cct //