LOCUS Exported 7437 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 pAAV-ProA5-GCaMP6s SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7437) 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 7437) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7437 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 2039..2048 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2045..3397 /codon_start=1 /product="improved fluorescent protein-based calcium sensor (Chen et al., 2013)" /label=GCaMP6s /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" regulatory 2141..2150 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 2431..2452 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2558..2579) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2819..2838) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 3434..4022 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3487..3507) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3905..3916) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4054..4530 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4253..4272) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4570..4710 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4570..4699 /label=AAV2 ITR rep_origin 4785..5240 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4872..4891) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5082..5103 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5257..5276) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5376..5398 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5436..5454) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5522..5626 /gene="bla" /label=AmpR promoter CDS 5627..6487 /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(5845..5864) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6658..7246 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7147..7166 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 7308..7437 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 gcggccgcac gcgtggtgcc caggcagtgg gagcagggct gaccagagtt ctgcagagat 61 tgcctggagg ccttcctgga agaagagatc ctggcaccgc acaaagagaa gcacaggctt 121 tccagggctg aggagaggga ggtcaagtga ggcccaggtg cccctgcctg agcctgtgtc 181 cccagaaacc tcctctccct ctcatcaccc ccacatcctc cctgccactc cccgcagctc 241 cctgtggcca agtgcactgc agcactcggc tctgctccac aaacggtctg ctccactcca 301 ggaaggccac ctcctccccc cccccccacc tccggctgtc accactcacc gctctagcct 361 ccagggggtg gggaccccag agctggacac accccatcga agccccacag ctcagccagc 421 cggacagact cacggtcgga ctcaagaccc cggagccctg aggtgggcag cgcgccaggg 481 ttcctcgcag cctcttcaag gtcagtgcaa gttgggtgtg cagccctttg gagcctttga 541 ggtctgtggg actcagctca gaaccctcat cccagaccac gagccccaag gtgggcttca 601 ctccaggtgc acctgagcca agtgctgggc aaagggcgcc caggtccaca gtcagcacag 661 gtccaggggt ccatcaggag gggccccagg ctgggggcca accccttgct gaggcctctt 721 tggggacact ccccaccggc catggggagc taaaattgga aagtggcgag tgggaggcag 781 ctgacacagc tattttgtgc gctcttcatc cgctccggtt tcactctcct gctccactaa 841 cctgatcctg tgcttccctc ccccaccctg ccgatgaaga ggtgtctcag ccctccagcc 901 acagtgcaga ggctgctcag gagggccagg ccgggagcga gcagggccag cgtggtctgg 961 tctgagccgg gactcaccag gagcacctgc ccttcagaga agctcactgg atggcctccc 1021 gggtccctga ccctttgtga cccacacctc tctgtcagtg tagcaaggag cggcctccca 1081 gagccccaac tacagggctg gaggagacag atctgcaact gggagatctt gggttgggag 1141 agatggggtg accctaccca tctccaaaag ggcactgcac cgtgttcccc tttccctcgg 1201 ttccaggtct ggacctaagc aaccactgga agactggcgg ccaggctgaa gagggtggga 1261 ggggcttagg tactggccca gaccccaggg atggcccagc agggtagcca gagggagtga 1321 ccagacacat cctggacttg ataccagtca gcaccttgga cagcagccag caccctccca 1381 gttgctgcct ggccctgcct ggtccagctc tggccactct ggggcctcaa agagcccaac 1441 atccagcctg cagcaagagc ttagactaca cagcacttca agggggacag tgggtgaaca 1501 caggcaggga cctgagataa aacagactca cggcttggcg gaggaatgga gtcagcagag 1561 cccggagttg agcgagtctg ggaagttagt gctagaggag ttcacgggat ctcaggggac 1621 ctcgtgggca gggctttgct gaggttaaga gaggctcact gtgagacggg agaactgaca 1681 gagtcttcca ccatgggaaa gaccccatgg ttggggaagt cttgcaggaa cagtttggtg 1741 tcccaaactt gtcagcccat cagagaggta tgaaggagag gaagctagtc tgtgcgaagt 1801 ctgggctttg gagaatcttc aaaggaagca agattgagag ctgcagagga aaggaagaag 1861 gtgtctgggt gtgttggggg gggtagattt actgacagca tccccagggg atggctgagt 1921 ggaccagtct cctcagaggg tccaccaggg gaggcgaggc aaggctggga ctgggggttc 1981 tcactcatgc ttctccttca gccctctcca ggccggatcc tgtacaccgc gggctagcgc 2041 caccatgggt tctcatcatc atcatcatca tggtatggcc agcatgactg gtggacagca 2101 aatgggtcgg gatctgtacg acgatgacga taaggatctc gccaccatgg tggactcatc 2161 acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact 2221 cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttccacat 2281 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc 2341 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact 2401 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 2461 cgggatcact ctcggcatgg acgagctgta taagggcggt accggaggga gcatggtgag 2521 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 2581 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct 2641 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 2701 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 2761 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga 2821 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 2881 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 2941 gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc 3001 cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg 3061 gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc 3121 cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaaatgaa 3181 atacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata aggatggcaa 3241 tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac 3301 agatgaagag gttgatgaaa tgatcaggga agcagacatc gacggggatg gtcaggtaaa 3361 ctacgaagag tttgtgcaaa tgatgacagc gaagtgagtc gacagatctg aattcgatat 3421 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 3481 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 3541 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 3601 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 3661 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 3721 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 3781 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 3841 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 3901 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 3961 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 4021 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca 4081 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa 4141 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg 4201 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg 4261 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt 4321 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg 4381 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc 4441 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag 4501 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg 4561 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctccgcgc gctcgctcgc 4621 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag 4681 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc 4741 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg 4801 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 4861 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 4921 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 4981 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac 5041 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 5101 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat 5161 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 5221 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata 5281 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 5341 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 5401 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 5461 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 5521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 5581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 5641 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 5701 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 5761 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 5821 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 5881 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 5941 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 6001 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 6061 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 6121 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 6181 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 6241 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 6301 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 6361 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 6421 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 6481 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 6541 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 6601 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 6661 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 6721 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 6781 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 6841 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 6901 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 6961 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 7021 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 7081 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 7141 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 7201 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 7261 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct 7321 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 7381 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcct //