LOCUS Exported 6551 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 pAAV2-ProC3-Cre/mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6551) 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 6551) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6551 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 57..66 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 63..1091 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" CDS 1101..1154 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 1155..1862 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(1297..1315) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1565..1584 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1890..2478 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1943..1963) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2361..2372) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2510..2986 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2709..2728) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3026..3166 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3026..3155 /label=AAV2 ITR rep_origin 3241..3696 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3328..3347) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3538..3559 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3713..3732) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3832..3854 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3892..3910) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3978..4082 /gene="bla" /label=AmpR promoter CDS 4083..4943 /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(4301..4320) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5114..5702 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5603..5622 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 5764..5893 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 6405..50 /label=SV40 promoter /note="SV40 early promoter" rep_origin 6452..36 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6514..6533 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa ggatccgcca 61 ccatggccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga 121 gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc 181 atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata 241 accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg 301 cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc 361 gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgaa 421 tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca 481 ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta 541 atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga 601 tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa 661 cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac 721 tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt 781 gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg 841 ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg 901 gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg 961 cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata 1021 ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg 1081 aagatggcga tggaccggtc gagggcagag gaagtctgct aacatgcggt gacgtcgagg 1141 agaatcctgg cccagtgagc aagggcgagg aggataacat ggccatcatc aaggagttca 1201 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 1261 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg 1321 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct 1381 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca 1441 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 1501 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg 1561 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc 1621 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc 1681 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg 1741 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg 1801 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt 1861 aatgagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 1921 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 1981 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 2041 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 2101 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 2161 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 2221 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 2281 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga 2341 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 2401 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 2461 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc 2521 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 2581 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 2641 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 2701 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 2761 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 2821 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 2881 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 2941 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 3001 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 3061 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 3121 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 3181 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 3241 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3301 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3361 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3421 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 3481 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3541 gactcttgtt ccaaactgga acaacactca actctatttc gggctattct tttgatttat 3601 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 3661 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 3721 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 3781 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 3841 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 3901 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 3961 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 4021 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 4081 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 4141 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 4201 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 4261 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 4321 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 4381 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 4441 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 4501 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 4561 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 4621 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 4681 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 4741 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 4801 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 4861 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 4921 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 4981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 5041 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 5101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 5161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 5221 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 5281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 5341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 5461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 5521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 5581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 5641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 5701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 5761 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc 5821 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 5881 actaggggtt cctgcggccg cacgcgtaaa gctggaactg gggccggaaa cagctgaggg 5941 tgcccagccg gaaactcgaa atcaacgtag gccggaaact attcgatgaa ttccgccgga 6001 aactaggtcc ggaggactgc cggaaacacc tgtaatcaag ccgccggaaa cctgttgtgg 6061 ccgtatgccg gaaacgtctt aattggacgt gccggaaact cttttaatga gttcgccgga 6121 aaccagacca gccgagctgc cggaaaccgg ttatatagaa cggccggaaa cggtccacag 6181 gaaaaagccg gaaacaccca aacggttagc gccggaaacg actggggagg acgtgccgga 6241 aacgtactat ctgaagatgc cggaaacact tgaaagctcc aagccggaaa cgggcccgtg 6301 cggatagccg gaaactgacg gtacacggcc gccggaaaca ctacttgtat ggtagccgga 6361 aacttgggcg tggctggggc cggaaacgct cgagatctgc gatctgcatc tcaattagtc 6421 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 6481 ccattctccg ccccatcgct gactaatttt ttttatttat gcagaggccg aggccgcctc 6541 ggcctctgag c //