LOCUS       Exported                6825 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Targeting expression to neuronal and glial cell types in mice, 
            non-human primates, and human retina.
ACCESSION   .
VERSION     .
KEYWORDS    IV_pAAV-ProB6-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  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 6825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6825
                     /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      1269..1278
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1275..2198
                     /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
                     RKTTKLNIGGTEIEVETLVEDEAEAGA"
     CDS             2205..2921
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2250..2271)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2511..2530)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2858..2879
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2952..3540
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3005..3025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3423..3434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3572..4048
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3771..3790)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4088..4228
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4088..4217
                     /label=AAV2 ITR
     rep_origin      4303..4758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4390..4409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4600..4621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4775..4794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4894..4916
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4954..4972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5040..5144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5145..6005
                     /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(5363..5382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6176..6764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6665..6684
                     /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 gcgtgttaag aatctttctc taatgttggg cttaagtggc
      181 aggagggata tatatcagga tgcaattcct tgcatctggt ttgtaatctg cttgggttgc
      241 ttgaggaaat gcacttaatt aaaataaaca ttaaacatcc tagcaacaga ggacttatag
      301 gctaataagg ttgaggattt tgtgatttaa agaactcctt aatccactgc ttcctgccta
      361 ccataagcca aggggctgtt caaagattct cttaaacaat tcaactcaaa aattattcac
      421 agttcataaa cagagtgcaa atcacccaag cagaagtaat ttactgcttt taagtcaaag
      481 ccctgcttag ctagggagtc agctgtagga attaaccaga acaaaatcct gattaaacag
      541 ctaatcggtt gttctttggg ttccatattt tcccttcaag tctagctgga ttgcaaagtt
      601 aagcttatga ttatgaataa aaactatttt ccacaggttt ttctttaatc atttgtaagg
      661 aaatcaatct caatgcccta ttattgtctt tttttctgaa cctgaaaaac cctctctttt
      721 ctcccaggcc ccctactccg ctaacccagt ggcctgagca aatcacaagg aggattggtg
      781 gctaaataag aaggcagcag ctttggattt tccactcatg gaagagggga ggaaagggca
      841 gagggtgggg gcttccttgg gaagcttggg atgagtcacg ggagtggagg gtggatagga
      901 tatggagagg ggcggggccg cagccctaac ccaaagtgtt gtcacaccaa acccttcaca
      961 cctggccttt gtctccccct ggcggggcac cgccctgaat gaccactgtg taatttttat
     1021 ttagaatttc gctctgaagt catcagctct atgacccaga aacactgaat aataaaagac
     1081 cctgccaact ggaatgtgat accctgatct ctggcctttg gaggtggtat ttagggcaat
     1141 tgagtatatt ttcaaggaaa tagtgactta taccagcaag ttgtcttact acaaatattt
     1201 gtaagtaaac accaccctaa gtacagcttc ctgctctttc atccctttta aggtcattgc
     1261 caggatccgc caccatggat tatggaggcg ccctgagtgc cgttgggcgc gagctgctat
     1321 ttgtaacgaa cccagtagtc gtcaatggct ctgtacttgt gcctgaggac cagtgttact
     1381 gcgcgggctg gattgagtcg cgtggcacaa acggtgccca aacggcgtcg aacgtgctgc
     1441 aatggcttgc tgctggcttc tccatcctac tgcttatgtt ttacgcctac caaacatgga
     1501 agtcaacctg cggctgggag gagatctatg tgtgcgctat cgagatggtc aaggtgattc
     1561 tcgagttctt cttcgagttt aagaacccgt ccatgctgta tctagccaca ggccaccgcg
     1621 tccagtggtt gcgttacgcc gagtggcttc tcacctgccc ggtcatttgc attcacctgt
     1681 caaacctgac gggcttgtcc aacgactaca gcaggcgcac catgggtctg cttgtgtctg
     1741 atattggcac aattgtgtgg ggcgccactt ccgccatggc caccggatac gtcaaggtca
     1801 tcttcttctg cctgggtctg tgttatggtg ctaacacgtt ctttcacgct gccaaggcct
     1861 acatcgaggg ttaccacacc gtgccgaagg gccggtgtcg ccaggtggtg actggcatgg
     1921 cttggctctt cttcgtatca tggggtatgt tccccatcct gttcatcctc ggccccgagg
     1981 gcttcggcgt cctgagcgtg tacggctcca ccgtcggcca caccatcatt gacctgatgt
     2041 cgaagaactg ctggggtctg ctcggccact acctgcgcgt gctgatccac gagcatatcc
     2101 tcatccacgg cgacattcgc aagaccacca aattgaacat tggtggcact gagattgagg
     2161 tcgagacgct ggtggaggac gaggccgagg ctggcgcggt acccatggtg agcaagggcg
     2221 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2281 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2341 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2401 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2461 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2521 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2581 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2641 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2701 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2761 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2821 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2881 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgataagaa ttcgatatca
     2941 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3001 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3061 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3121 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3181 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3241 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3301 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3361 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3421 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3481 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3541 atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     3601 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     3661 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     3721 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     3781 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     3841 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     3901 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     3961 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     4021 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     4081 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4141 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4201 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4261 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4321 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     4381 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4441 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4501 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4561 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4621 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4681 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4741 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4801 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4861 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4921 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4981 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
