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" promoter 661..692 /label=minP /note="minimal TATA-box promoter with low basal activity" regulatory 746..755 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 752..1675 /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 1682..2398 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1727..1748) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1988..2007) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2335..2356 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2429..3017 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2482..2502) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2900..2911) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3049..3525 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3248..3267) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3565..3705 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3565..3694 /label=AAV2 ITR rep_origin 3780..4235 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3867..3886) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4077..4098 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4252..4271) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4371..4393 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4431..4449) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4517..4621 /gene="bla" /label=AmpR promoter CDS 4622..5482 /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(4840..4859) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5653..6241 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6142..6161 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 6303..6432 /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 atcagataga gttgaacaag 121 cagtaccaaa gcgggtagca ttggtgccgc gtgcggtaac tcagtgtaac tcgttcaccg 181 gcagacttca acattcattt ctgactgatt cagcagtcag agcgccaaac ttccagtaat 241 ctctctgaag atttggcaca gatacggagt tcagaatagg aacaagtgtg cactgatgga 301 gaggagctac tacggctatt tctaaaaata gcatggctgc aggcttccgg aggcttagga 361 ttctcacgaa acgaaactga gagattctta agatagaacc aggagactgt agctgaagat 421 tcctgactga cgtgaatatg acttccttta tcccaaagga tgcagtgact tctcaggctc 481 gggtagcatc atgattagtt ataattgact ttgcctttta atttttaagg caatctataa 541 attatggatt gttgtgtcaa tcggaagaca ggattttgac atcgtacacc atatacctca 601 tgattgtcgt caaaaccaag tcctttatcg gggatcctca catggtcctg ctggagttag 661 tagagggtat ataatggaag ctcgacttcc agctatcaca tccactgtgt tgttgtgaac 721 tggaatccac tataggccag gatccgccac catggattat ggaggcgccc tgagtgccgt 781 tgggcgcgag ctgctatttg taacgaaccc agtagtcgtc aatggctctg tacttgtgcc 841 tgaggaccag tgttactgcg cgggctggat tgagtcgcgt ggcacaaacg gtgcccaaac 901 ggcgtcgaac gtgctgcaat ggcttgctgc tggcttctcc atcctactgc ttatgtttta 961 cgcctaccaa acatggaagt caacctgcgg ctgggaggag atctatgtgt gcgctatcga 1021 gatggtcaag gtgattctcg agttcttctt cgagtttaag aacccgtcca tgctgtatct 1081 agccacaggc caccgcgtcc agtggttgcg ttacgccgag tggcttctca cctgcccggt 1141 catttgcatt cacctgtcaa acctgacggg cttgtccaac gactacagca ggcgcaccat 1201 gggtctgctt gtgtctgata ttggcacaat tgtgtggggc gccacttccg ccatggccac 1261 cggatacgtc aaggtcatct tcttctgcct gggtctgtgt tatggtgcta acacgttctt 1321 tcacgctgcc aaggcctaca tcgagggtta ccacaccgtg ccgaagggcc ggtgtcgcca 1381 ggtggtgact ggcatggctt ggctcttctt cgtatcatgg ggtatgttcc ccatcctgtt 1441 catcctcggc cccgagggct tcggcgtcct gagcgtgtac ggctccaccg tcggccacac 1501 catcattgac ctgatgtcga agaactgctg gggtctgctc ggccactacc tgcgcgtgct 1561 gatccacgag catatcctca tccacggcga cattcgcaag accaccaaat tgaacattgg 1621 tggcactgag attgaggtcg agacgctggt ggaggacgag gccgaggctg gcgcggtacc 1681 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 1741 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 1801 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 1861 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 1921 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 1981 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 2041 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 2101 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 2161 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 2221 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 2281 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 2341 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg 2401 ataagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag 2461 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 2521 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 2581 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 2641 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 2701 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 2761 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 2821 gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac 2881 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 2941 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 3001 ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc 3061 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc 3121 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt 3181 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc 3241 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc 3301 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag 3361 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata 3421 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa 3481 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta 3541 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc 3601 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg 3661 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt 3721 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta 3781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3841 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 3961 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc 4021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 4081 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata 4141 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa 4201 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg 4261 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg 4321 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg 4381 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat 4441 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac 4501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 4561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 4621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 4681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 4741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 4801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 4861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 4921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 4981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 5041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 5101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 5161 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 5221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 5281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 5341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 5401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 5461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 5521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 5581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 5641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 5701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 5761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 5821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 5881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 5941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 6001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 6061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 6121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 6181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 6241 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 6301 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct 6361 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca 6421 ctaggggttc ct //