LOCUS       Exported                7707 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    127_pAAV-ProA32-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  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 7707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7707
                     /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      2151..2160
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2157..3080
                     /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             3087..3803
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3132..3153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3393..3412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3740..3761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3834..4422
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3887..3907)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4305..4316)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4454..4930
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4653..4672)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4970..5110
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4970..5099
                     /label=AAV2 ITR
     rep_origin      5185..5640
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5272..5291)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5482..5503
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5657..5676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5776..5798
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5836..5854)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5922..6026
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6027..6887
                     /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(6245..6264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7058..7646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7547..7566
                     /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 gcgtacttga ggccaccctg gtctctatat cgtgagttcc
      181 agctagccag tgctacatag tgagccccta cttcaaagac aaaatagaat tagagactaa
      241 gttgacagct cagctggtag tgcttggtta tcatgcagcc ttggaccaac ttcagccctg
      301 catgcactgg gtatgaaagc taacacctga aattcctcat tggagaggtg gaggcaggag
      361 aatgaaggct atcctcggct acagagtgac ctcaaggtca gattggaata taagagatcc
      421 tgtctcaaaa acaaagcaaa caagcaacaa cggaaagaat aaaagtaaga aaaatattga
      481 ctgaaaagct ttgagcttga tgaaagattc tcaatcagca aatttagaaa gcccaaagag
      541 ttcaaagtac aacaaacgca gagacagtgt ggcaaaacac agagcagagt gggcgcacgc
      601 ctttaatccc agcactcggg aggcagaggc aggcagattt ctgagttcga agccagcctg
      661 gtctacagag tgagttccag gacagtcagg gctacacaga gaaaccctgt ctcaaaaaac
      721 caaaaaacca aaaccaaaaa caaaacaaaa caaaacaaaa ctagactgag ggaaacctga
      781 gtttccttta aaagcacaag gcgtataatg gagcagtggg tgactatgat gaactgaagc
      841 cctaaaatct caagaaagca gaacaaaatg tcataaaagc tagactttca tagattcagt
      901 acacatgaag cagtagtagc agaacacaag gaaagaacag atggagagga aaacaatcct
      961 gaagagcaag aaatgagagg ccaaaagaaa cagtacatct aatgggaaaa ccaacgtaaa
     1021 gcagcagaaa taaacccagg tacaccaaca aatacaataa gtataaatgg attagatgca
     1081 caagctaaaa gacaaagact accccttgga aacttttatt atgaaaataa cttcaggtgt
     1141 atagaacatg ggaagtacat gaacaacatg gatattcctg ccatatagag tcatgtgtac
     1201 tcttcacacg aatgtttgat gaggaggcta tgaaggtcta ttttacatag tatacgttac
     1261 acaatgaaat actagtcacg tgccaatggg aacatacagg gcttattttg ggtatcagac
     1321 agagctaggc tctcagttac ttttagtcat gtgatttatg tgtgtaacta atttattctc
     1381 tcagtgaggt cctagcacgg tgtgaagcag gtcgttttct ggttattgta tttctaaata
     1441 tttcttgcat aaacaagtaa tgctatcagc aaatggttca gtgcagcgct ttgcggtttg
     1501 gtaaacactg aatgcagaat agcttttatt tatgacagtt actgaaagca ttcagcggag
     1561 tggcacgaaa taagcaaaca ataattgcca gtttattata tttataattg ttatttatga
     1621 actgtcacaa ggcctggcac atatcactaa cccgtaaaca ttgtgtttgt aagaataata
     1681 agtactttat tatattcagg ctcagtagag cctggcacca cacgtgcggg aagaaagtgg
     1741 aagtttattc ttgttaatta ttattattaa gtaaagcact taacaacagt tctccccgca
     1801 ggcgcttact gaacagtctc actggagacc gcccctcctc ccaagcctcg agaggccacg
     1861 ccccctgtct cggaagctcc gccctctacc gcccggtctg tgcctgcgca gtggccctga
     1921 acaccctgcg catgcgcagt ctcccttttg tcgctggagc cgctggccac tgccggagcc
     1981 cggcatctgc tacgtacgtg cgtacgtgcg tgcgtcgggg cgtgcgtggg tacgtcgtgc
     2041 gtgccgtgag ctcgcccggc accgcggcct cgccctccgc ccctgcgccc cgccacccag
     2101 gccgccgccc cccgcccgcg ccgtcgcccc cgacggggag gaccggatcc gccaccatgg
     2161 attatggagg cgccctgagt gccgttgggc gcgagctgct atttgtaacg aacccagtag
     2221 tcgtcaatgg ctctgtactt gtgcctgagg accagtgtta ctgcgcgggc tggattgagt
     2281 cgcgtggcac aaacggtgcc caaacggcgt cgaacgtgct gcaatggctt gctgctggct
     2341 tctccatcct actgcttatg ttttacgcct accaaacatg gaagtcaacc tgcggctggg
     2401 aggagatcta tgtgtgcgct atcgagatgg tcaaggtgat tctcgagttc ttcttcgagt
     2461 ttaagaaccc gtccatgctg tatctagcca caggccaccg cgtccagtgg ttgcgttacg
     2521 ccgagtggct tctcacctgc ccggtcattt gcattcacct gtcaaacctg acgggcttgt
     2581 ccaacgacta cagcaggcgc accatgggtc tgcttgtgtc tgatattggc acaattgtgt
     2641 ggggcgccac ttccgccatg gccaccggat acgtcaaggt catcttcttc tgcctgggtc
     2701 tgtgttatgg tgctaacacg ttctttcacg ctgccaaggc ctacatcgag ggttaccaca
     2761 ccgtgccgaa gggccggtgt cgccaggtgg tgactggcat ggcttggctc ttcttcgtat
     2821 catggggtat gttccccatc ctgttcatcc tcggccccga gggcttcggc gtcctgagcg
     2881 tgtacggctc caccgtcggc cacaccatca ttgacctgat gtcgaagaac tgctggggtc
     2941 tgctcggcca ctacctgcgc gtgctgatcc acgagcatat cctcatccac ggcgacattc
     3001 gcaagaccac caaattgaac attggtggca ctgagattga ggtcgagacg ctggtggagg
     3061 acgaggccga ggctggcgcg gtacccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3121 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3181 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3241 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3301 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3361 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3421 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3481 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3661 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3781 tcggcatgga cgagctgtac aagtgataag aattcgatat caagcttatc gataatcaac
     3841 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3901 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3961 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4021 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4081 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4141 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4201 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     4261 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4321 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4381 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     4441 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     4501 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     4561 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     4621 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     4681 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     4741 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     4801 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     4861 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     4921 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     4981 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     5041 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     5101 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5161 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5221 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     5281 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5341 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5401 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5461 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     5521 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     5581 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5641 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5701 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5761 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5821 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5881 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5941 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6001 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6061 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6121 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6181 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6241 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6301 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6361 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6421 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6481 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6541 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6601 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6661 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6721 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6781 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6841 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6901 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7201 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7681 ccttttgctg gccttttgct cacatgt
//
