LOCUS       sequence-241142-        6432 bp    DNA     circular SYN 15-APR-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 6432)
  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 6432)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: 185_pAAV-ProD16-CatCh-GFP-WPRE
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        791..822
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      876..885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             882..1808
                     /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
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             join(1812..1814,1815..1817,1818..2531)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1812..1814,1815..1817,1818..2528)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1857..1878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2118..2137)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2559..3147
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2612..2632)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3030..3041)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3179..3655
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3695..3835
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3695..3824
                     /label=AAV2 ITR
     rep_origin      3910..4365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3997..4016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4207..4228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4382..4401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4501..4523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4561..4579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4647..4751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4752..4820,4821..5612)
                     /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(4970..4989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5783..6371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6272..6291
                     /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 gcgtatcctg gggtttaaac aggactaggt ccactgggag
      181 aagaggaagt caaacaggac tagtcatcga taggctagcc agcttaatta atcagataga
      241 gttgaacaag cagtaccaaa gcgggtagca ttggtgccgc gtgcggtaac tcagtgtaac
      301 tcgttcaccg gcagacttca acattcattt ctgactgatt cagcagtcag agcgccaaac
      361 ttccagtaat ctctctgaag atttggcaca gatacggagt tcagaatagg aacaagtgtg
      421 cactgatgga gaggagctac tacggctatt tctaaaaata gcatggctgc aggcttccgg
      481 aggcttagga ttctcacgaa acgaaactga gagattctta agatagaacc aggagactgt
      541 agctgaagat tcctgactga cgtgaatatg acttccttta tcccaaagga tgcagtgact
      601 tctcaggctc gggtagcatc atgattagtt ataattgact ttgcctttta atttttaagg
      661 caatctataa attatggatt gttgtgtcaa tcggaagaca ggattttgac atcgtacacc
      721 atatacctca tgattgtcgt caaaaccaag tcctttatcg gggatcctca catggtcctg
      781 ctggagttag tagagggtat ataatggaag ctcgacttcc agctatcaca tccactgtgt
      841 tgttgtgaac tggaatccac tataggccaa gatccgccac catggattat ggaggcgccc
      901 tgagtgccgt tgggcgcgag ctgctatttg taacgaaccc agtagtcgtc aatggctctg
      961 tacttgtgcc tgaggaccag tgttactgcg cgggctggat tgagtcgcgt ggcacaaacg
     1021 gtgcccaaac ggcgtcgaac gtgctgcaat ggcttgctgc tggcttctcc atcctactgc
     1081 ttatgtttta cgcctaccaa acatggaagt caacctgcgg ctgggaggag atctatgtgt
     1141 gcgctatcga gatggtcaag gtgattctcg agttcttctt cgagtttaag aacccgtcca
     1201 tgctgtatct agccacaggc caccgcgtcc agtggttgcg ttacgccgag tggcttctca
     1261 cctgcccggt catttgcatt cacctgtcaa acctgacggg cttgtccaac gactacagca
     1321 ggcgcaccat gggtctgctt gtgtctgata ttggcacaat tgtgtggggc gccacttccg
     1381 ccatggccac cggatacgtc aaggtcatct tcttctgcct gggtctgtgt tatggtgcta
     1441 acacgttctt tcacgctgcc aaggcctaca tcgagggtta ccacaccgtg ccgaagggcc
     1501 ggtgtcgcca ggtggtgact ggcatggctt ggctcttctt cgtatcatgg ggtatgttcc
     1561 ccatcctgtt catcctcggc cccgagggct tcggcgtcct gagcgtgtac ggctccaccg
     1621 tcggccacac catcattgac ctgatgtcga agaactgctg gggtctgctc ggccactacc
     1681 tgcgcgtgct gatccacgag catatcctca tccacggcga cattcgcaag accaccaaat
     1741 tgaacattgg tggcactgag attgaggtcg agacgctggt ggaggacgag gccgaggctg
     1801 gcgcggtacc catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1861 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1921 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1981 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2041 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2101 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2161 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2221 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2281 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2341 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2401 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2461 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2521 tgtacaagtg ataagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2581 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2641 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2701 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2761 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2821 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2881 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2941 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     3001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3061 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3121 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     3181 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3241 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3301 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3361 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3421 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     3481 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     3541 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3601 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3661 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3721 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3781 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3841 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3901 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4321 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4381 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4561 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6421 ttgctcacat gt
//
