LOCUS       Exported                7348 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    IX_pAAV-ProB3-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7348)
  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 7348)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7348
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        212..514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      1792..1801
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1798..2721
                     /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             2728..3444
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2773..2794)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3034..3053)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3381..3402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3475..4063
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3528..3548)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3946..3957)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4095..4571
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4294..4313)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4611..4751
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4611..4740
                     /label=AAV2 ITR
     rep_origin      4826..5281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4913..4932)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5123..5144
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5298..5317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5417..5439
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5477..5495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5563..5667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5668..6528
                     /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(5886..5905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6699..7287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7188..7207
                     /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 gcgtctagca ctagttatta atagtaatca attacggggt
      181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      301 taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc
      361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      481 agtacatcta cgtattagtc atcgctatta ccatcactct agtgttttca tgtgacctct
      541 tgttctcaat ttagatactt cttgacctgg aaaaggtggg tgctgcttgc cactctacat
      601 ttggctactc ctgttttagg atgtgcatgt tccttcctcc ccttgtctcc cgccatgaaa
      661 tctcttccca tcctctttgc aacctgacaa cagagccttg tccctaggta gatatacaca
      721 tacattgttt ttctcattgt gttccttatt tttagagaga gaaaaaaaat ctgagtctgt
      781 aagataaatg caaaggataa caatccccaa aacagcctag gaacccctca ttcctctggc
      841 aagacactac tgatgatctg tatgcttggt agcctcttta agtcctctca gagatagtat
      901 agtagttgac aaatgccagc tcctatattc tctagcttct ccttttcctc tatcaaatga
      961 ctttttaaat aaattagata ttagacaaac cttgggtgac ctgctttttc ttttcttttc
     1021 cttttttttt ttttacttct ccccccgccc cccctccccc ccgttttttt tcccttgcgg
     1081 tataatagtg ttgaagaaag agggcactag tgtacagccc agatggcagc cttgcaccgt
     1141 ctgatttgga gctgaggtgt ctgcgaaccg agggcggctg cagtcctctg acctgagacc
     1201 aatagctctg tctacccgga ctcaggtagg tcttgaagat gaaagggtgt gtgtgtgtgt
     1261 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt acgcgcgcgc gcgcgcgagt
     1321 gtacgaatcc tagctctgag cgctgtccag ggttctgaca tttgagtggc tcaggcggtt
     1381 caagccccaa cacagggagg ctaccaggcg ttgccttagt aactaatatg gcttttccac
     1441 aagctgtcgc atttacattt tccaccatca gcgtccagct tatggccact aacgcgccgc
     1501 tcattgcaca cctgattcac acaccttcgg gcaccagtga aaaaaccgcg acttgatttt
     1561 ctggaagaac acccctaggg tttgggagcg gtcgtggagg accagcagga agaagcggag
     1621 gggagagggg cggtggtgga gactgagaag ctttgaacca gctgtgttgg ccaaaggcac
     1681 gaagcggcaa aagacagcgg cgggcgtcag ggtggttcgc gctgggaacc tgcaggcggg
     1741 accggcgtgg gaacgtggct tgccggcggt cgaaccgcgt cttcgggatc cgccaccatg
     1801 gattatggag gcgccctgag tgccgttggg cgcgagctgc tatttgtaac gaacccagta
     1861 gtcgtcaatg gctctgtact tgtgcctgag gaccagtgtt actgcgcggg ctggattgag
     1921 tcgcgtggca caaacggtgc ccaaacggcg tcgaacgtgc tgcaatggct tgctgctggc
     1981 ttctccatcc tactgcttat gttttacgcc taccaaacat ggaagtcaac ctgcggctgg
     2041 gaggagatct atgtgtgcgc tatcgagatg gtcaaggtga ttctcgagtt cttcttcgag
     2101 tttaagaacc cgtccatgct gtatctagcc acaggccacc gcgtccagtg gttgcgttac
     2161 gccgagtggc ttctcacctg cccggtcatt tgcattcacc tgtcaaacct gacgggcttg
     2221 tccaacgact acagcaggcg caccatgggt ctgcttgtgt ctgatattgg cacaattgtg
     2281 tggggcgcca cttccgccat ggccaccgga tacgtcaagg tcatcttctt ctgcctgggt
     2341 ctgtgttatg gtgctaacac gttctttcac gctgccaagg cctacatcga gggttaccac
     2401 accgtgccga agggccggtg tcgccaggtg gtgactggca tggcttggct cttcttcgta
     2461 tcatggggta tgttccccat cctgttcatc ctcggccccg agggcttcgg cgtcctgagc
     2521 gtgtacggct ccaccgtcgg ccacaccatc attgacctga tgtcgaagaa ctgctggggt
     2581 ctgctcggcc actacctgcg cgtgctgatc cacgagcata tcctcatcca cggcgacatt
     2641 cgcaagacca ccaaattgaa cattggtggc actgagattg aggtcgagac gctggtggag
     2701 gacgaggccg aggctggcgc ggtacccatg gtgagcaagg gcgaggagct gttcaccggg
     2761 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2821 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2881 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     2941 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3001 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3061 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3121 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3181 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3241 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3301 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     3361 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3421 ctcggcatgg acgagctgta caagtgataa gaattcgata tcaagcttat cgataatcaa
     3481 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3541 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3601 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3661 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3721 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3781 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3841 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     3901 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3961 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4021 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4081 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4141 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4201 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     4261 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     4321 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     4381 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4441 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     4501 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     4561 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     4621 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4681 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4741 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4801 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4861 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4921 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4981 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5041 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5101 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     5161 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     5221 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5281 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5341 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5401 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5461 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5521 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5581 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5641 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5701 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5761 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5821 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5881 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5941 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6001 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6061 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6121 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6181 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6241 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6301 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6361 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6421 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6481 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6541 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6601 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6841 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7321 gccttttgct ggccttttgc tcacatgt
//
