LOCUS       Exported                6332 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    210_pAAV-ProD19-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6332)
  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 6332)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6332
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        561..592
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      646..655
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             652..1575
                     /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             1582..2298
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1627..1648)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1888..1907)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2235..2256
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2329..2917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2382..2402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2800..2811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2949..3425
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3465..3605
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3465..3594
                     /label=AAV2 ITR
     rep_origin      3680..4135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3767..3786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3977..3998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4152..4171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4271..4293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4331..4349)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4417..4521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4522..5382
                     /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(4740..4759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5553..6141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6042..6061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6203..6332
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcggccgcac gcgtatcctg gggtttaaac aggactaggt ccactgggag aagaggaagt
       61 caaacaggac tagtcatcga taggctagcc agcttaatta atcagatgag accgcagagc
      121 tacagttcag ttgcagcttg agtctgcctc cctgatagag aaacggaatc aagaccagct
      181 gtgagacgga ggctgaaaga cacagagcac ttcagagagt gagcggggct gcgggtagtc
      241 tgggacaggg gagcctcctt ggggcaagcc cagcaggggg tgtttgggaa agaaagttca
      301 ccaggagttg gagtatgact ctctgtcctg gttcagccct gtgtggccgt gtgacacgga
      361 cagattggct gtcacttgga tgtggcagga tcatagctgc tgagtggagc agagggtggg
      421 agggtgcatt gggctgcacc gtggagggag gagaggagag caaacagact gggcagaagt
      481 gaggcaggtc agagatgggt ctgctgatgg catcttcctc aggcacttta aggatcctca
      541 catggtcctg ctggagttag tagagggtat ataatggaag ctcgacttcc agctatcaca
      601 tccactgtgt tgttgtgaac tggaatccac tataggccag gatccgccac catggattat
      661 ggaggcgccc tgagtgccgt tgggcgcgag ctgctatttg taacgaaccc agtagtcgtc
      721 aatggctctg tacttgtgcc tgaggaccag tgttactgcg cgggctggat tgagtcgcgt
      781 ggcacaaacg gtgcccaaac ggcgtcgaac gtgctgcaat ggcttgctgc tggcttctcc
      841 atcctactgc ttatgtttta cgcctaccaa acatggaagt caacctgcgg ctgggaggag
      901 atctatgtgt gcgctatcga gatggtcaag gtgattctcg agttcttctt cgagtttaag
      961 aacccgtcca tgctgtatct agccacaggc caccgcgtcc agtggttgcg ttacgccgag
     1021 tggcttctca cctgcccggt catttgcatt cacctgtcaa acctgacggg cttgtccaac
     1081 gactacagca ggcgcaccat gggtctgctt gtgtctgata ttggcacaat tgtgtggggc
     1141 gccacttccg ccatggccac cggatacgtc aaggtcatct tcttctgcct gggtctgtgt
     1201 tatggtgcta acacgttctt tcacgctgcc aaggcctaca tcgagggtta ccacaccgtg
     1261 ccgaagggcc ggtgtcgcca ggtggtgact ggcatggctt ggctcttctt cgtatcatgg
     1321 ggtatgttcc ccatcctgtt catcctcggc cccgagggct tcggcgtcct gagcgtgtac
     1381 ggctccaccg tcggccacac catcattgac ctgatgtcga agaactgctg gggtctgctc
     1441 ggccactacc tgcgcgtgct gatccacgag catatcctca tccacggcga cattcgcaag
     1501 accaccaaat tgaacattgg tggcactgag attgaggtcg agacgctggt ggaggacgag
     1561 gccgaggctg gcgcggtacc catggtgagc aagggcgagg agctgttcac cggggtggtg
     1621 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1681 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1741 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1801 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1861 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1921 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1981 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2041 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2101 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2161 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2221 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2281 atggacgagc tgtacaagtg ataagaattc gatatcaagc ttatcgataa tcaacctctg
     2341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2461 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2521 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2581 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2641 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2701 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2761 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2821 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2881 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     2941 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3001 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3061 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3121 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3181 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3241 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3301 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3361 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3421 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3481 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3541 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3601 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3661 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3721 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     3781 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3841 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3901 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3961 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4021 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4081 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4141 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4201 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4261 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4321 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4381 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4501 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5341 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6181 tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     6241 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     6301 gggagtggcc aactccatca ctaggggttc ct
//
