LOCUS Exported 6299 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 132_pAAV-ProB2-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6299) 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 6299) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6299 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 743..752 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 749..1672 /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 1679..2395 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1724..1745) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1985..2004) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2332..2353 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2426..3014 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2479..2499) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2897..2908) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3046..3522 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3245..3264) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3562..3702 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3562..3691 /label=AAV2 ITR rep_origin 3777..4232 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3864..3883) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4074..4095 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4249..4268) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4368..4390 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4428..4446) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4514..4618 /gene="bla" /label=AmpR promoter CDS 4619..5479 /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(4837..4856) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5650..6238 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6139..6158 /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 gcgtgaccat ttaaaagggg tatataaaga ttgcatacaa 181 aagctgaggg gcctcaccct gaatggattc tttcttgaaa gccacttttg ttctttaaga 241 ggcgcggtcc aaataatgtg caagcatgat tggctggagg cacatttcgt agattaatcc 301 tctcccttga aaggatccaa gtctggaaaa tagccaaaaa cgtggctgtt atgcaaccct 361 tacccaagct cctcctccca gctgccatcc ctcttactaa gcagtgtcat gaggctgggc 421 caggctggag attaattctt gaccatttaa aaggggtata taaagattgc atacaaaagc 481 tgaggggcct caccctgaat ggattctttc ttgaaagcca cttttgttct ttaagaggcg 541 cggtccaaat aatgtgcaag catgattggc tggaggcaca tttcgtagat taatcctctc 601 ccttgaaagg atccaagtct ggaaaatagc caaaaacgtg gctgttatgc aacccttacc 661 caagctcctc ctcccagctg ccatccctct tactaagcag tgtcatgagg ctgggccagg 721 ctggagatta attcttagat ccgccaccat ggattatgga ggcgccctga gtgccgttgg 781 gcgcgagctg ctatttgtaa cgaacccagt agtcgtcaat ggctctgtac ttgtgcctga 841 ggaccagtgt tactgcgcgg gctggattga gtcgcgtggc acaaacggtg cccaaacggc 901 gtcgaacgtg ctgcaatggc ttgctgctgg cttctccatc ctactgctta tgttttacgc 961 ctaccaaaca tggaagtcaa cctgcggctg ggaggagatc tatgtgtgcg ctatcgagat 1021 ggtcaaggtg attctcgagt tcttcttcga gtttaagaac ccgtccatgc tgtatctagc 1081 cacaggccac cgcgtccagt ggttgcgtta cgccgagtgg cttctcacct gcccggtcat 1141 ttgcattcac ctgtcaaacc tgacgggctt gtccaacgac tacagcaggc gcaccatggg 1201 tctgcttgtg tctgatattg gcacaattgt gtggggcgcc acttccgcca tggccaccgg 1261 atacgtcaag gtcatcttct tctgcctggg tctgtgttat ggtgctaaca cgttctttca 1321 cgctgccaag gcctacatcg agggttacca caccgtgccg aagggccggt gtcgccaggt 1381 ggtgactggc atggcttggc tcttcttcgt atcatggggt atgttcccca tcctgttcat 1441 cctcggcccc gagggcttcg gcgtcctgag cgtgtacggc tccaccgtcg gccacaccat 1501 cattgacctg atgtcgaaga actgctgggg tctgctcggc cactacctgc gcgtgctgat 1561 ccacgagcat atcctcatcc acggcgacat tcgcaagacc accaaattga acattggtgg 1621 cactgagatt gaggtcgaga cgctggtgga ggacgaggcc gaggctggcg cggtacccat 1681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 1801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 1861 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 2101 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg 2161 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga 2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgata 2401 agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt 2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 2581 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 2641 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 2701 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 2761 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 2821 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc 2881 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 2941 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 3001 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt 3061 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt 3121 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg 3181 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc 3241 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc 3301 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca 3361 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg 3421 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg 3481 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc 3541 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc 3601 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc 3661 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt 3721 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc 3781 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3841 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3901 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3961 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc 4021 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 4081 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg 4141 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 4201 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc 4261 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 4321 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 4381 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg 4441 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 4501 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 4561 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 4621 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 4681 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 4741 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 4801 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 4861 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 4921 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 4981 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 5041 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 5101 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 5161 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 5221 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 5281 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 5341 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 5401 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 5461 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 5521 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 5581 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 5641 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 5701 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 5761 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 5821 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 5881 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 5941 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 6001 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 6061 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 6121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 6181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 6241 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt //