LOCUS       Exported                7024 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Targeting expression to neuronal and glial cell types in mice, 
            non-human primates, and human retina.
ACCESSION   .
VERSION     .
KEYWORDS    III_pAAV-ProB4-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7024)
  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 7024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7024
                     /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      1468..1477
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1474..2397
                     /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             2404..3120
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2449..2470)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2710..2729)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3057..3078
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3151..3739
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3204..3224)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3622..3633)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3771..4247
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3970..3989)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4287..4427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4287..4416
                     /label=AAV2 ITR
     rep_origin      4502..4957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4589..4608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4799..4820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4974..4993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5093..5115
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5153..5171)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5239..5343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5344..6204
                     /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(5562..5581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6375..6963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6864..6883
                     /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 gcgtgttctt tgggttccat attttcctca ttccccttca
      181 agtctagctg gattgcaaag ttaagcttat gattatgaat aaaaactatt gagtatattt
      241 tcaaggaaat agtgacttac cttttaaggt cacttcctgc tctttcatct gacccagaaa
      301 cactgaataa taaagccatc atttgttgaa tgaccactgt gtaattttta tttagaactc
      361 taatgttggt aatctgcttg gggctgtggg ttaagaatct ttgggttgct tgaggaaatg
      421 cacttaatta aaataaacat taaacttttc cacactggcc tttgtctccc cctggcggat
      481 cctagcaaca gataccagca agttgtctta cttggtattt agggcaatgt gatgcagccc
      541 taacccaaag tgttgtcaca ccaaaccctt cacacccaag aggatgtttc gctctgaagt
      601 catcagctct aacaaatatt tgtaagtaaa caggtttttc tttaaggagg ccttcatttg
      661 taaccaacac catgctctcc tttttcgctc tctgaagtca tcagctctaa aggaaatcaa
      721 tctcaatgcc ctattatacc ctgatctctg gcctttggag gggctgttca aagattctct
      781 gaggacttat aggctaataa ggttgaggat tttgtgattt aaagaactcc ttaatccact
      841 gcttcctgcc taccataagc caaggccacc cttgcctttt tttttcttat cccctgctga
      901 acctgaaaaa ccctctcttt tctacagttt tctgttccca ggccccaagt acagttaaac
      961 aattcaactc aacttgttta atcctctcgt cttaaattat tcacattgcc aagtatttca
     1021 acagaatgta ctggcaagtt cataaacaga gtgcaaatca cccaagcaga agtaatttac
     1081 tgcttttaag tcaaagccct gcttagctag ggagtcagct gtaggaatta accagaacaa
     1141 aatcctgatt aaacagctaa tcggttctac tccgctaacc cagtggcctg agcaaatcac
     1201 aaggaggatt ggttccttgc atctggtttg ggcaccgccc acctggaacc ctaggggcgg
     1261 ggccgcttaa gtggcaggag ggatatatat caggatgcaa aagaccctgc caactggaaa
     1321 aataagaagg cagcagcttt ggattttcca ctcatggaag aggggaggaa agggcagagg
     1381 gtgggggctt ccttgggaag cttgggatga gtcaccacgg gattggggag tggagggtgg
     1441 ataggatatg gagaagggtg gagatccgcc accatggatt atggaggcgc cctgagtgcc
     1501 gttgggcgcg agctgctatt tgtaacgaac ccagtagtcg tcaatggctc tgtacttgtg
     1561 cctgaggacc agtgttactg cgcgggctgg attgagtcgc gtggcacaaa cggtgcccaa
     1621 acggcgtcga acgtgctgca atggcttgct gctggcttct ccatcctact gcttatgttt
     1681 tacgcctacc aaacatggaa gtcaacctgc ggctgggagg agatctatgt gtgcgctatc
     1741 gagatggtca aggtgattct cgagttcttc ttcgagttta agaacccgtc catgctgtat
     1801 ctagccacag gccaccgcgt ccagtggttg cgttacgccg agtggcttct cacctgcccg
     1861 gtcatttgca ttcacctgtc aaacctgacg ggcttgtcca acgactacag caggcgcacc
     1921 atgggtctgc ttgtgtctga tattggcaca attgtgtggg gcgccacttc cgccatggcc
     1981 accggatacg tcaaggtcat cttcttctgc ctgggtctgt gttatggtgc taacacgttc
     2041 tttcacgctg ccaaggccta catcgagggt taccacaccg tgccgaaggg ccggtgtcgc
     2101 caggtggtga ctggcatggc ttggctcttc ttcgtatcat ggggtatgtt ccccatcctg
     2161 ttcatcctcg gccccgaggg cttcggcgtc ctgagcgtgt acggctccac cgtcggccac
     2221 accatcattg acctgatgtc gaagaactgc tggggtctgc tcggccacta cctgcgcgtg
     2281 ctgatccacg agcatatcct catccacggc gacattcgca agaccaccaa attgaacatt
     2341 ggtggcactg agattgaggt cgagacgctg gtggaggacg aggccgaggc tggcgcggta
     2401 cccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2461 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2521 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2581 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2641 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2701 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2761 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2821 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2881 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2941 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3001 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3061 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3121 tgataagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     3181 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3241 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3301 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3361 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3421 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3481 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3541 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
     3601 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3661 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3721 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
     3781 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3841 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3901 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3961 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     4021 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     4081 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     4141 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     4201 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     4261 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4321 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4381 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4441 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4501 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4561 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4621 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4681 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4741 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4801 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     4861 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     4921 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4981 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5041 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5101 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5161 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5221 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5281 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5341 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5401 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5461 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5521 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5581 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5641 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5701 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5761 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5821 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5881 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5941 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6001 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6061 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6121 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6181 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6241 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6301 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6361 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6421 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6481 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6541 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6601 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6661 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6721 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6781 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6841 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6901 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6961 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7021 atgt
//
