LOCUS Exported 6730 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 83_pAAV-ProC14-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6730) 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 6730) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6730 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 841..1037 /label=SV40 promoter /note="SV40 early promoter" rep_origin 888..1023 /label=SV40 ori /note="SV40 origin of replication" primer_bind 950..969 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" regulatory 1044..1053 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1050..1973 /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 1980..2696 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2025..2046) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2286..2305) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2633..2654 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2727..3315 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2780..2800) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3198..3209) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3347..3823 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3546..3565) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3863..4003 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3863..3992 /label=AAV2 ITR rep_origin 4078..4533 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4165..4184) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4375..4396 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4550..4569) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4669..4691 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4729..4747) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4815..4919 /gene="bla" /label=AmpR promoter CDS 4920..5780 /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(5138..5157) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5951..6539 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6440..6459 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 6601..6730 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 gcggccgcac gcgttctaac tcgctgtagt aattccagcg agaggcagag ggagcgagcg 61 ggcggccacg cgttcaagcc tcctaccgcc tttgttatgc aaacatatca aacgccctcc 121 tttgttatgc aaaagggctg gaacggggcc tttgttatgc aaatcgccct ccccgatccc 181 tttgttatgc aaatttgacg aattcccacc tttgttatgc aaacaaatct cctaccctcc 241 tttgttatgc aaagtgagag gggctgcacc tttgttatgc aaaatgcggc ccctgagacc 301 tttgttatgc aaaaaccatg tacgcttgcc tttgttatgc aaaccgcctg ttgcttggcc 361 tttgttatgc aaagccacgc gattggcgcc tttgttatgc aaagctcggt tatgtacacc 421 tttgttatgc aaagctactt taaacttgcc tttgttatgc aaatcacgac ctgaccgtcc 481 tttgttatgc aaaaacggtt gaaatagtcc tttgttatgc aaaatgatat tgaatagtcc 541 tttgttatgc aaaaatttag atgccgaccc tttgttatgc aaaggaatgg gcgtgctgcc 601 tttgttatgc aaattttcgc tgcgacagcc tttgttatgc aaacatgctc gccactcacc 661 tttgttatgc aaaggtctaa caatgacccc tttgttatgc aaactacgtg gaatagatcc 721 tttgttatgc aaaccccgag tttttgaacc tttgttatgc aaaatcagta acttcattcc 781 tttgttatgc aaaatgtgac ttaacctccc tttgttatgc aaagctcgag atctgcgatc 841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 901 ctccgcccag ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag 961 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 1021 gcctaggctt ttgcaaagga tccgccacca tggattatgg aggcgccctg agtgccgttg 1081 ggcgcgagct gctatttgta acgaacccag tagtcgtcaa tggctctgta cttgtgcctg 1141 aggaccagtg ttactgcgcg ggctggattg agtcgcgtgg cacaaacggt gcccaaacgg 1201 cgtcgaacgt gctgcaatgg cttgctgctg gcttctccat cctactgctt atgttttacg 1261 cctaccaaac atggaagtca acctgcggct gggaggagat ctatgtgtgc gctatcgaga 1321 tggtcaaggt gattctcgag ttcttcttcg agtttaagaa cccgtccatg ctgtatctag 1381 ccacaggcca ccgcgtccag tggttgcgtt acgccgagtg gcttctcacc tgcccggtca 1441 tttgcattca cctgtcaaac ctgacgggct tgtccaacga ctacagcagg cgcaccatgg 1501 gtctgcttgt gtctgatatt ggcacaattg tgtggggcgc cacttccgcc atggccaccg 1561 gatacgtcaa ggtcatcttc ttctgcctgg gtctgtgtta tggtgctaac acgttctttc 1621 acgctgccaa ggcctacatc gagggttacc acaccgtgcc gaagggccgg tgtcgccagg 1681 tggtgactgg catggcttgg ctcttcttcg tatcatgggg tatgttcccc atcctgttca 1741 tcctcggccc cgagggcttc ggcgtcctga gcgtgtacgg ctccaccgtc ggccacacca 1801 tcattgacct gatgtcgaag aactgctggg gtctgctcgg ccactacctg cgcgtgctga 1861 tccacgagca tatcctcatc cacggcgaca ttcgcaagac caccaaattg aacattggtg 1921 gcactgagat tgaggtcgag acgctggtgg aggacgaggc cgaggctggc gcggtaccca 1981 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 2041 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 2101 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 2161 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 2221 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 2281 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 2341 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 2401 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 2461 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 2521 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 2581 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 2641 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgat 2701 aagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat 2761 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 2821 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 2881 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 2941 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 3001 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 3061 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 3121 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt 3181 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 3241 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 3301 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg 3361 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct 3421 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat 3481 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg 3541 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc 3601 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc 3661 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt 3721 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt 3781 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac 3841 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg 3901 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc 3961 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat 4021 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg 4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 4141 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 4261 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat 4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 4381 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag 4441 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg 4501 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct 4561 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac 4621 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca 4681 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac 4741 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt 4801 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 4861 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 4921 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 4981 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 5041 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 5101 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 5161 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 5221 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 5281 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 5341 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 5401 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 5461 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 5521 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 5581 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 5641 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 5701 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 5761 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 5821 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 5881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 6061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 6301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 6361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 6421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 6481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 6541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 6601 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 6661 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 6721 aggggttcct //