LOCUS       Exported                6325 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    II_pAAV-ProB5-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6325)
  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 6325)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6325
                     /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      769..778
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             775..1698
                     /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             1705..2421
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1750..1771)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2011..2030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2358..2379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2452..3040
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2505..2525)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2923..2934)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3072..3548
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3588..3728
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3588..3717
                     /label=AAV2 ITR
     rep_origin      3803..4258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3890..3909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4100..4121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4275..4294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4394..4416
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4454..4472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4540..4644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4645..5505
                     /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(4863..4882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5676..6264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6165..6184
                     /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 aaaaactaag ccagaagttg
      241 ctcacattta ggaggcttgt gtgctggggg ttgggaggat gactgctaaa tactggggaa
      301 gcaaaatgag ctaagtcttt acagcaactg tcatttgttt ttccacaggt ttttctttaa
      361 ggaggccttc atttgtaacc aacaccatgc tctccttaag gaaatcaatc tcaatgccct
      421 attatccttc ccttttcttt cctcccagtt tgaggctgca gttgcctttt tttttcttat
      481 cccctgctga acctgaaaaa ccctctcttt tctacagttt tctgttccca ggccccgctg
      541 acttccttta gagcatgggg ggggggggat caggattgtg atgtgtgaac tgggaggatc
      601 ttgacctact ccgctaaccc agtggcctga gcaaatcaca aggaggattg gagccatctg
      661 cccagcccct cccccacggc agcctgctgg aaagagacaa gttagtcatt caaatgattg
      721 gctttttgcc cgcttcttct ctaaataaga aggcagcagc ttggatccgc caccatggat
      781 tatggaggcg ccctgagtgc cgttgggcgc gagctgctat ttgtaacgaa cccagtagtc
      841 gtcaatggct ctgtacttgt gcctgaggac cagtgttact gcgcgggctg gattgagtcg
      901 cgtggcacaa acggtgccca aacggcgtcg aacgtgctgc aatggcttgc tgctggcttc
      961 tccatcctac tgcttatgtt ttacgcctac caaacatgga agtcaacctg cggctgggag
     1021 gagatctatg tgtgcgctat cgagatggtc aaggtgattc tcgagttctt cttcgagttt
     1081 aagaacccgt ccatgctgta tctagccaca ggccaccgcg tccagtggtt gcgttacgcc
     1141 gagtggcttc tcacctgccc ggtcatttgc attcacctgt caaacctgac gggcttgtcc
     1201 aacgactaca gcaggcgcac catgggtctg cttgtgtctg atattggcac aattgtgtgg
     1261 ggcgccactt ccgccatggc caccggatac gtcaaggtca tcttcttctg cctgggtctg
     1321 tgttatggtg ctaacacgtt ctttcacgct gccaaggcct acatcgaggg ttaccacacc
     1381 gtgccgaagg gccggtgtcg ccaggtggtg actggcatgg cttggctctt cttcgtatca
     1441 tggggtatgt tccccatcct gttcatcctc ggccccgagg gcttcggcgt cctgagcgtg
     1501 tacggctcca ccgtcggcca caccatcatt gacctgatgt cgaagaactg ctggggtctg
     1561 ctcggccact acctgcgcgt gctgatccac gagcatatcc tcatccacgg cgacattcgc
     1621 aagaccacca aattgaacat tggtggcact gagattgagg tcgagacgct ggtggaggac
     1681 gaggccgagg ctggcgcggt acccatggtg agcaagggcg aggagctgtt caccggggtg
     1741 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1801 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1861 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1921 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1981 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2041 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2101 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2161 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2221 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2281 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2341 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2401 ggcatggacg agctgtacaa gtgataagaa ttcgatatca agcttatcga taatcaacct
     2461 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2521 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2581 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2641 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2701 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2761 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2821 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     2881 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2941 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3001 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3061 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3121 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3181 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3241 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3301 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3361 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3421 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3481 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3541 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     3601 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3661 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3721 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3781 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3841 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     3901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3961 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4141 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4201 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4261 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4321 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4381 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4441 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4501 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4561 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4621 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4681 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4741 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4801 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4861 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4921 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4981 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5041 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5101 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5161 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5221 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5281 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5341 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5401 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5461 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5521 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5581 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5641 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5701 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5761 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5821 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5881 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5941 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6001 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6061 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6301 ttttgctggc cttttgctca catgt
//
