LOCUS Exported 6432 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 185_pAAV-ProD16-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6432) 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 6432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6432 /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 791..822 /label=minP /note="minimal TATA-box promoter with low basal activity" regulatory 876..885 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 882..1805 /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 1812..2528 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1857..1878) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2118..2137) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2465..2486 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2559..3147 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2612..2632) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3030..3041) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3179..3655 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3378..3397) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3695..3835 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3695..3824 /label=AAV2 ITR rep_origin 3910..4365 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3997..4016) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4207..4228 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4382..4401) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4501..4523 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4561..4579) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4647..4751 /gene="bla" /label=AmpR promoter CDS 4752..5612 /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(4970..4989) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5783..6371 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6272..6291 /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 atcagataga 241 gttgaacaag cagtaccaaa gcgggtagca ttggtgccgc gtgcggtaac tcagtgtaac 301 tcgttcaccg gcagacttca acattcattt ctgactgatt cagcagtcag agcgccaaac 361 ttccagtaat ctctctgaag atttggcaca gatacggagt tcagaatagg aacaagtgtg 421 cactgatgga gaggagctac tacggctatt tctaaaaata gcatggctgc aggcttccgg 481 aggcttagga ttctcacgaa acgaaactga gagattctta agatagaacc aggagactgt 541 agctgaagat tcctgactga cgtgaatatg acttccttta tcccaaagga tgcagtgact 601 tctcaggctc gggtagcatc atgattagtt ataattgact ttgcctttta atttttaagg 661 caatctataa attatggatt gttgtgtcaa tcggaagaca ggattttgac atcgtacacc 721 atatacctca tgattgtcgt caaaaccaag tcctttatcg gggatcctca catggtcctg 781 ctggagttag tagagggtat ataatggaag ctcgacttcc agctatcaca tccactgtgt 841 tgttgtgaac tggaatccac tataggccaa gatccgccac catggattat ggaggcgccc 901 tgagtgccgt tgggcgcgag ctgctatttg taacgaaccc agtagtcgtc aatggctctg 961 tacttgtgcc tgaggaccag tgttactgcg cgggctggat tgagtcgcgt ggcacaaacg 1021 gtgcccaaac ggcgtcgaac gtgctgcaat ggcttgctgc tggcttctcc atcctactgc 1081 ttatgtttta cgcctaccaa acatggaagt caacctgcgg ctgggaggag atctatgtgt 1141 gcgctatcga gatggtcaag gtgattctcg agttcttctt cgagtttaag aacccgtcca 1201 tgctgtatct agccacaggc caccgcgtcc agtggttgcg ttacgccgag tggcttctca 1261 cctgcccggt catttgcatt cacctgtcaa acctgacggg cttgtccaac gactacagca 1321 ggcgcaccat gggtctgctt gtgtctgata ttggcacaat tgtgtggggc gccacttccg 1381 ccatggccac cggatacgtc aaggtcatct tcttctgcct gggtctgtgt tatggtgcta 1441 acacgttctt tcacgctgcc aaggcctaca tcgagggtta ccacaccgtg ccgaagggcc 1501 ggtgtcgcca ggtggtgact ggcatggctt ggctcttctt cgtatcatgg ggtatgttcc 1561 ccatcctgtt catcctcggc cccgagggct tcggcgtcct gagcgtgtac ggctccaccg 1621 tcggccacac catcattgac ctgatgtcga agaactgctg gggtctgctc ggccactacc 1681 tgcgcgtgct gatccacgag catatcctca tccacggcga cattcgcaag accaccaaat 1741 tgaacattgg tggcactgag attgaggtcg agacgctggt ggaggacgag gccgaggctg 1801 gcgcggtacc catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 1861 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 1921 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 1981 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 2041 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 2101 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 2161 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 2221 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca 2281 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg 2341 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 2401 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg 2461 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 2521 tgtacaagtg ataagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa 2581 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 2641 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 2701 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 2761 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 2821 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 2881 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 2941 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc 3001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 3061 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 3121 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg 3181 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt 3241 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt 3301 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac 3361 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct 3421 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt 3481 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg 3541 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg 3601 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat 3661 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc 3721 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc 3781 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc 3841 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc 3901 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 4021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac 4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt 4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 4321 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca 4381 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg 4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg 4561 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 4621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6421 ttgctcacat gt //