LOCUS       sequence-256786-        7437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting expression to neuronal and glial cell types in mice, 
            non-human primates, and human retina.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Neurosci. 2019 Jul 8. pii: 10.1038/s41593-019-0431-2. doi: 
            10.1038/s41593-019-0431-2."
COMMENT     Sequence Label: pAAV-ProA5-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..7437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2039..2048
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2045..2053,2054..2071,2072..2074,2075..2107,2108..2110,
                     2111..2134,2135..2161,2162..2218,2219..2224,2225..2494,
                     2495..2512,2513..2947,2948..2953,2954..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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             join(5627..5695,5696..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
//
