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-ProA1-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" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 2169..2178 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2175..3527 /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 2271..2280 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 2561..2582 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2688..2709) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2949..2968) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 3564..4152 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3617..3637) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4035..4046) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4184..4660 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4383..4402) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4700..4840 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4700..4829 /label=AAV2 ITR rep_origin 4915..5370 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5002..5021) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5212..5233 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5387..5406) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5506..5528 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5566..5584) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5652..5756 /gene="bla" /label=AmpR promoter CDS 5757..6617 /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(5975..5994) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6788..7376 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7277..7296 /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 gcgtagaggc aggccgagtt tgaggccagc ctggtctaca 181 caggcgttct aggagaacct gtctcatatg cacatgggcc tgggattaca catacaactg 241 caatggcagc acttggaaag cggaagcagg agaatcggaa tttcacactc atccttcatt 301 atagaagtcc aaacctgtgc taagctactt ggacatcgct aaaaaacagc aaaaatcttt 361 ctaggaattg ccaatgtata cacaccaagt tgtatttttg tagggcagac ttttccaact 421 tgccagatga taataatcat ttatattagc tacatttcag gctccagaat taacactgtt 481 actgaaatgt ataggttcta aaacatacca tttccaaata ttttaaagga ttaacatttt 541 tgaaaagcat gtgatcttcc aaccatattt tagggaacag acataagaag taggcaagtt 601 tgggaagaaa cagttcctga gggcatttct tcccaacctt gaatgccctg tgggttaact 661 gaggtcttgg taaaatgtag attatttact ggactttttg aactgagagt gcatttctaa 721 caaaactcca gggtatgtgg tccttgcatt acatatgggg tagcaacatt ctaaagcagt 781 gtttttcaac ctgtggatca agacccacag aagtcacata gcagatatcc tgcctattag 841 atatttacat taagattcag agcagtagca gaataggagt catcccaacc tgaggaactg 901 catcagagtc ccagcatcag gaagggtgag agccactgag ctaaagtcct ctaggtgagg 961 gggctgcccc ggaaagactc atttaaatga aaccactgac acagagagct gacagatgag 1021 gtgggttccg tgtctgtgag gctctgctgg tcgtctcctc actcccatgg aagaaccacc 1081 gagatgaggg cgaggggcag agctaaccca gcctctaagt aggcagagtc tagtgtccag 1141 ctgcccaagg aagaagtttg ctgtgtgagg tggccctgat gtccgcacac acaaaatgcc 1201 agtgaagtct acttgaccaa gtgaagctgg tgtggaatgg gaagaagcac acacagtcag 1261 tctctctgca cacactctgt ccttcacttc ttcacttacc agagatttga tgagaaccta 1321 ctagcagatc agatttgatc cctgagtgga aaaaacgtac agtgggagat aaaagaggaa 1381 aacaacggat ctgagtttga ggttagccta gtctgagcaa gccggataca cagtaagacc 1441 ctgtctcaca tacatccact cgcacgcgcg cgcacacaaa cacacacaca cacacacaca 1501 cacacacaca cacacacaca catcgtgcta aggacaagat aggcatcctg agagatgagc 1561 caggacaaag aaccagtaat agctcctgga gcagcacatc tgttttgcca ggattatccc 1621 ttggatctct taaaaccgag accttgtaat ctgaagactc aacttgggct gtacccttaa 1681 ccttcagctc tatgatgcaa gtgagtccac aggaccggag gctttgagat gagcttttca 1741 gaagggagga gttggccgct tgctcccaga gctccagcac ctgcattctt ctggctatgt 1801 cagaagccag atcatttccc tcgttaaaaa caaaaacaaa aaaacaaaca aacaaaatgt 1861 tagtctttgc cctttatctg cctggcaaag cttttaattg gcttgatctg tcattccgct 1921 agacataaag gggacaatcc ccggattagg aaggagctct ccagctcggg taaggagtct 1981 caaggcaagg taggcaagca ccaccggtcc gcactctcgc ccagctttta cgggaagaag 2041 agaatgttac tctatcctaa catatttttc cttttcctct atctcacaga taggaaaaat 2101 ttaagagcca gagggaacgt cccttctcag aggagacagc agaaggatcc tgtacaccgc 2161 gggctagcgc caccatgggt tctcatcatc atcatcatca tggtatggcc agcatgactg 2221 gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatctc gccaccatgg 2281 tggactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc 2341 tgagctcact cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga 2401 acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc 2461 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc 2521 agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 2581 tgaccgccgc cgggatcact ctcggcatgg acgagctgta taagggcggt accggaggga 2641 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct 2761 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 2821 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct 2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 3061 acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag 3121 aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga 3181 cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg 3241 aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa 3301 gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata 3361 aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag 3421 agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc gacggggatg 3481 gtcaggtaaa ctacgaagag tttgtgcaaa tgatgacagc gaagtgagtc gacagatctg 3541 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga 3601 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 3661 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 3721 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 3781 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 3841 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 3901 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat 3961 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct 4021 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 4081 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 4141 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga 4201 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt 4261 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg 4321 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg 4381 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc 4441 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg 4501 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc 4561 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct 4621 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac 4681 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc 4741 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg 4801 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt 4861 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc 4921 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 4981 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 5041 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 5101 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc 5161 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 5221 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga 5281 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 5341 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg 5401 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 5461 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 5521 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc 5581 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc 5641 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 5701 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 5761 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 5821 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 5881 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 5941 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 6001 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 6061 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 6121 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 6181 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 6241 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 6301 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 6361 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 6421 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 6481 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 6541 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 6601 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 6661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 6721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 6781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 6841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6901 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 6961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 7021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 7081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 7141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 7201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 7261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 7321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 7381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt //