LOCUS       Exported                6468 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    177_pAAV-ProD8-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6468)
  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 6468)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6468
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        827..858
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      912..921
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             918..1841
                     /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             1848..2564
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1893..1914)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2154..2173)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2501..2522
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2595..3183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2648..2668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3066..3077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3215..3691
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3731..3871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3731..3860
                     /label=AAV2 ITR
     rep_origin      3946..4401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4033..4052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4243..4264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4418..4437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4537..4559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4597..4615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4683..4787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4788..5648
                     /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(5006..5025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5819..6407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6308..6327
                     /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 atcagatcgg
      241 gagggttcaa acgagcgtga gcatccaaac cgtacagctg aaggtctcct acactcagac
      301 agccagcaac cggcacttgc tctgtgcctg tagacacttc ctgtctcata tccaattcac
      361 ctctataaac ctcatcctcc cgggccccca aagcctccac gggatgtctt aaaacgagaa
      421 gagccgcttc ctgaacctct tttcagtggc tctgtcttta aagtctcagc tctgggatgg
      481 cttcctccta agctcctgta ttcaaatccc caggctagca caaagtatct ctccctgtgt
      541 acttctctaa tctaataaat ctctttcatc ttgacatgaa gtattagaag tcattttcag
      601 acttgtaaag atgttggtta cttaagacac gcattaattg attagcacat gacagagttt
      661 cctgatactg aggatcgatg tggaaagaaa gatacttagt tactctaagc ctctatctaa
      721 ggagacaggg aggaaacgtt acttttcata cataggtcag acatgaatct aactagggga
      781 tggttggttt tgtttggcaa tcctcacatg gtcctgctgg agttagtaga gggtatataa
      841 tggaagctcg acttccagct atcacatcca ctgtgttgtt gtgaactgga atccactata
      901 ggccaggatc cgccaccatg gattatggag gcgccctgag tgccgttggg cgcgagctgc
      961 tatttgtaac gaacccagta gtcgtcaatg gctctgtact tgtgcctgag gaccagtgtt
     1021 actgcgcggg ctggattgag tcgcgtggca caaacggtgc ccaaacggcg tcgaacgtgc
     1081 tgcaatggct tgctgctggc ttctccatcc tactgcttat gttttacgcc taccaaacat
     1141 ggaagtcaac ctgcggctgg gaggagatct atgtgtgcgc tatcgagatg gtcaaggtga
     1201 ttctcgagtt cttcttcgag tttaagaacc cgtccatgct gtatctagcc acaggccacc
     1261 gcgtccagtg gttgcgttac gccgagtggc ttctcacctg cccggtcatt tgcattcacc
     1321 tgtcaaacct gacgggcttg tccaacgact acagcaggcg caccatgggt ctgcttgtgt
     1381 ctgatattgg cacaattgtg tggggcgcca cttccgccat ggccaccgga tacgtcaagg
     1441 tcatcttctt ctgcctgggt ctgtgttatg gtgctaacac gttctttcac gctgccaagg
     1501 cctacatcga gggttaccac accgtgccga agggccggtg tcgccaggtg gtgactggca
     1561 tggcttggct cttcttcgta tcatggggta tgttccccat cctgttcatc ctcggccccg
     1621 agggcttcgg cgtcctgagc gtgtacggct ccaccgtcgg ccacaccatc attgacctga
     1681 tgtcgaagaa ctgctggggt ctgctcggcc actacctgcg cgtgctgatc cacgagcata
     1741 tcctcatcca cggcgacatt cgcaagacca ccaaattgaa cattggtggc actgagattg
     1801 aggtcgagac gctggtggag gacgaggccg aggctggcgc ggtacccatg gtgagcaagg
     1861 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1921 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1981 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2041 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2101 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2161 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2221 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2281 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2341 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2401 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2461 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2521 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgataa gaattcgata
     2581 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2641 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2701 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2761 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2821 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2881 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2941 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3001 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3061 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3121 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3181 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     3241 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3301 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3361 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3421 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3481 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3541 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     3601 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     3661 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3721 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3781 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3841 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3901 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4021 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4141 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4201 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4261 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     4321 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4381 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4441 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4501 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4561 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4621 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4681 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4741 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4801 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4861 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4921 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4981 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5041 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5101 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5161 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5221 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5281 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5341 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5401 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5461 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5521 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5581 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5641 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5701 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5761 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5821 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5881 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5941 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6001 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6061 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6121 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6181 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6241 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6301 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6361 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6421 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
