LOCUS       sequence-257168-        6299 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Targeting expression to neuronal and glial cell types in mice, 
            non-human primates, and human retina.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  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 6299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Neurosci. 2019 Jul 8. pii: 10.1038/s41593-019-0431-2. doi: 
            10.1038/s41593-019-0431-2."
COMMENT     Sequence Label: 132_pAAV-ProB2-CatCh-GFP-WPRE
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      743..752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             749..1675
                     /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
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             join(1679..1681,1682..1684,1685..2398)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1679..1681,1682..1684,1685..2395)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1724..1745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1985..2004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2332..2353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2426..3014
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2479..2499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2897..2908)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3046..3522
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3245..3264)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3562..3702
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3562..3691
                     /label=AAV2 ITR
     rep_origin      3777..4232
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3864..3883)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4074..4095
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4249..4268)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4368..4390
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4428..4446)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4514..4618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4619..4687,4688..5479)
                     /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(4837..4856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5650..6238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6139..6158
                     /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 gcgtgaccat ttaaaagggg tatataaaga ttgcatacaa
      181 aagctgaggg gcctcaccct gaatggattc tttcttgaaa gccacttttg ttctttaaga
      241 ggcgcggtcc aaataatgtg caagcatgat tggctggagg cacatttcgt agattaatcc
      301 tctcccttga aaggatccaa gtctggaaaa tagccaaaaa cgtggctgtt atgcaaccct
      361 tacccaagct cctcctccca gctgccatcc ctcttactaa gcagtgtcat gaggctgggc
      421 caggctggag attaattctt gaccatttaa aaggggtata taaagattgc atacaaaagc
      481 tgaggggcct caccctgaat ggattctttc ttgaaagcca cttttgttct ttaagaggcg
      541 cggtccaaat aatgtgcaag catgattggc tggaggcaca tttcgtagat taatcctctc
      601 ccttgaaagg atccaagtct ggaaaatagc caaaaacgtg gctgttatgc aacccttacc
      661 caagctcctc ctcccagctg ccatccctct tactaagcag tgtcatgagg ctgggccagg
      721 ctggagatta attcttagat ccgccaccat ggattatgga ggcgccctga gtgccgttgg
      781 gcgcgagctg ctatttgtaa cgaacccagt agtcgtcaat ggctctgtac ttgtgcctga
      841 ggaccagtgt tactgcgcgg gctggattga gtcgcgtggc acaaacggtg cccaaacggc
      901 gtcgaacgtg ctgcaatggc ttgctgctgg cttctccatc ctactgctta tgttttacgc
      961 ctaccaaaca tggaagtcaa cctgcggctg ggaggagatc tatgtgtgcg ctatcgagat
     1021 ggtcaaggtg attctcgagt tcttcttcga gtttaagaac ccgtccatgc tgtatctagc
     1081 cacaggccac cgcgtccagt ggttgcgtta cgccgagtgg cttctcacct gcccggtcat
     1141 ttgcattcac ctgtcaaacc tgacgggctt gtccaacgac tacagcaggc gcaccatggg
     1201 tctgcttgtg tctgatattg gcacaattgt gtggggcgcc acttccgcca tggccaccgg
     1261 atacgtcaag gtcatcttct tctgcctggg tctgtgttat ggtgctaaca cgttctttca
     1321 cgctgccaag gcctacatcg agggttacca caccgtgccg aagggccggt gtcgccaggt
     1381 ggtgactggc atggcttggc tcttcttcgt atcatggggt atgttcccca tcctgttcat
     1441 cctcggcccc gagggcttcg gcgtcctgag cgtgtacggc tccaccgtcg gccacaccat
     1501 cattgacctg atgtcgaaga actgctgggg tctgctcggc cactacctgc gcgtgctgat
     1561 ccacgagcat atcctcatcc acggcgacat tcgcaagacc accaaattga acattggtgg
     1621 cactgagatt gaggtcgaga cgctggtgga ggacgaggcc gaggctggcg cggtacccat
     1681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1861 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2101 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2161 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgata
     2401 agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2581 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2641 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2701 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2761 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2821 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     2881 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2941 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3001 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     3061 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3121 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     3181 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     3241 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     3301 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     3361 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     3421 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     3481 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     3541 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     3601 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3661 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     3721 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     3781 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3841 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3901 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3961 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4021 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4081 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     4141 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4201 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     4261 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     4321 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4381 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     4441 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4501 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4561 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4621 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4681 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4741 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4801 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4861 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4921 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4981 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5041 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5101 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5161 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5221 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5281 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5341 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5401 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5461 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5521 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5581 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5641 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5701 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5761 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5821 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5881 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5941 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6001 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6061 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6241 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
