LOCUS       Exported                6259 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    160_pAAV-ProD4-CatCh-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  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 6259)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6259
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        618..649
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      703..712
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             709..1632
                     /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             1639..2355
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1684..1705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1945..1964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2292..2313
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2386..2974
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2439..2459)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2857..2868)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3006..3482
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3205..3224)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3522..3662
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3522..3651
                     /label=AAV2 ITR
     rep_origin      3737..4192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3824..3843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4034..4055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4209..4228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4328..4350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4388..4406)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4474..4578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4579..5439
                     /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(4797..4816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5610..6198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6099..6118
                     /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 gcgtatcctg gggtttaaac aggactaggt ccactgggag
      181 aagaggaagt caaacaggac tagtcatcga taggctagcc agcttaatta atcagatctc
      241 cagtgtttac ccagggcaag aagtcccatt tttgttcttt tacaagctga agaccagaaa
      301 cacatgcggc ccattcccgt ccgaggcaga aatgctgagg gataagtctc aggctctgag
      361 ctgagttcag gaaaaggtca gtgtcccgat agagcactca ggtagagtga atccgcttat
      421 cacagagtga atccagctta gttgtccttt ggaacctcag ttgattggat aagtcctcca
      481 agtgtgaaag aggcatctcc actcaggcct tggttccaat ggtctctcag ggaactgccc
      541 tcagcagtgc atagagctga gccagctctc cgggaagttc agtcgagttt atcctcacat
      601 ggtcctgctg gagttagtag agggtatata atggaagctc gacttccagc tatcacatcc
      661 actgtgttgt tgtgaactgg aatccactat aggccaggat ccgccaccat ggattatgga
      721 ggcgccctga gtgccgttgg gcgcgagctg ctatttgtaa cgaacccagt agtcgtcaat
      781 ggctctgtac ttgtgcctga ggaccagtgt tactgcgcgg gctggattga gtcgcgtggc
      841 acaaacggtg cccaaacggc gtcgaacgtg ctgcaatggc ttgctgctgg cttctccatc
      901 ctactgctta tgttttacgc ctaccaaaca tggaagtcaa cctgcggctg ggaggagatc
      961 tatgtgtgcg ctatcgagat ggtcaaggtg attctcgagt tcttcttcga gtttaagaac
     1021 ccgtccatgc tgtatctagc cacaggccac cgcgtccagt ggttgcgtta cgccgagtgg
     1081 cttctcacct gcccggtcat ttgcattcac ctgtcaaacc tgacgggctt gtccaacgac
     1141 tacagcaggc gcaccatggg tctgcttgtg tctgatattg gcacaattgt gtggggcgcc
     1201 acttccgcca tggccaccgg atacgtcaag gtcatcttct tctgcctggg tctgtgttat
     1261 ggtgctaaca cgttctttca cgctgccaag gcctacatcg agggttacca caccgtgccg
     1321 aagggccggt gtcgccaggt ggtgactggc atggcttggc tcttcttcgt atcatggggt
     1381 atgttcccca tcctgttcat cctcggcccc gagggcttcg gcgtcctgag cgtgtacggc
     1441 tccaccgtcg gccacaccat cattgacctg atgtcgaaga actgctgggg tctgctcggc
     1501 cactacctgc gcgtgctgat ccacgagcat atcctcatcc acggcgacat tcgcaagacc
     1561 accaaattga acattggtgg cactgagatt gaggtcgaga cgctggtgga ggacgaggcc
     1621 gaggctggcg cggtacccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1681 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1741 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1801 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1861 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1921 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1981 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2041 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2101 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2161 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2221 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2281 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2341 gacgagctgt acaagtgata agaattcgat atcaagctta tcgataatca acctctggat
     2401 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2461 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2521 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2581 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2641 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2701 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2761 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2821 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2881 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2941 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     3001 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     3061 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     3121 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     3181 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3241 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3301 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     3361 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     3421 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     3481 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3541 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3601 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3661 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3721 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3781 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3841 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3901 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3961 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4021 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     4081 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4141 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4201 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4261 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4321 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4381 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4441 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4501 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4561 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4621 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5401 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5461 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6241 tggccttttg ctcacatgt
//
