LOCUS       Exported                6167 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    157_pAAV-ProD2-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6167)
  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 6167)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6167
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        396..427
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      481..490
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             487..1410
                     /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             1417..2133
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1462..1483)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1723..1742)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2070..2091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2164..2752
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2217..2237)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2635..2646)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2784..3260
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2983..3002)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3300..3440
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3300..3429
                     /label=AAV2 ITR
     rep_origin      3515..3970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3602..3621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3812..3833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3987..4006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4106..4128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4166..4184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4252..4356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4357..5217
                     /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(4575..4594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5388..5976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5877..5896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6038..6167
                     /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 atcagatccc ttccagcatg
      121 acaataccag gttttcaagg gccctagaac tatgagatgg taaagtgatg cccccagatg
      181 tcctactcac agcacatgca gctgcttctc tctcccggct ctcccctcaa gaggcagcta
      241 tgcacaggaa tagtctaata atcccaaagt cacctaaata ctaatacaat taagactgta
      301 acagttaatc ccagacagca ctctgggcgg ggctagtctc gtacatggaa aggtcagtga
      361 aagcctctat cctcacatgg tcctgctgga gttagtagag ggtatataat ggaagctcga
      421 cttccagcta tcacatccac tgtgttgttg tgaactggaa tccactatag gccaggatcc
      481 gccaccatgg attatggagg cgccctgagt gccgttgggc gcgagctgct atttgtaacg
      541 aacccagtag tcgtcaatgg ctctgtactt gtgcctgagg accagtgtta ctgcgcgggc
      601 tggattgagt cgcgtggcac aaacggtgcc caaacggcgt cgaacgtgct gcaatggctt
      661 gctgctggct tctccatcct actgcttatg ttttacgcct accaaacatg gaagtcaacc
      721 tgcggctggg aggagatcta tgtgtgcgct atcgagatgg tcaaggtgat tctcgagttc
      781 ttcttcgagt ttaagaaccc gtccatgctg tatctagcca caggccaccg cgtccagtgg
      841 ttgcgttacg ccgagtggct tctcacctgc ccggtcattt gcattcacct gtcaaacctg
      901 acgggcttgt ccaacgacta cagcaggcgc accatgggtc tgcttgtgtc tgatattggc
      961 acaattgtgt ggggcgccac ttccgccatg gccaccggat acgtcaaggt catcttcttc
     1021 tgcctgggtc tgtgttatgg tgctaacacg ttctttcacg ctgccaaggc ctacatcgag
     1081 ggttaccaca ccgtgccgaa gggccggtgt cgccaggtgg tgactggcat ggcttggctc
     1141 ttcttcgtat catggggtat gttccccatc ctgttcatcc tcggccccga gggcttcggc
     1201 gtcctgagcg tgtacggctc caccgtcggc cacaccatca ttgacctgat gtcgaagaac
     1261 tgctggggtc tgctcggcca ctacctgcgc gtgctgatcc acgagcatat cctcatccac
     1321 ggcgacattc gcaagaccac caaattgaac attggtggca ctgagattga ggtcgagacg
     1381 ctggtggagg acgaggccga ggctggcgcg gtacccatgg tgagcaaggg cgaggagctg
     1441 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1501 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1561 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1621 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1681 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1741 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1801 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1861 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1921 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1981 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2041 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2101 gggatcactc tcggcatgga cgagctgtac aagtgataag aattcgatat caagcttatc
     2161 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2221 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2281 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2341 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2401 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2461 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2521 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2581 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2641 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2701 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     2761 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     2821 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     2881 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     2941 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3001 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3061 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3121 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     3181 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3241 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     3301 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3361 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3421 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3481 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     3541 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     3601 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3661 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3721 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3781 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3841 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     3901 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3961 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4021 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4081 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4141 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4201 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4261 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4321 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4381 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4441 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4501 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4561 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4621 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4681 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4741 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4801 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4861 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4921 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4981 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5041 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5101 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5161 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5221 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5521 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6001 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
     6061 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     6121 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcct
//
