LOCUS       sequence-137748-        8342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid can be used in the triple transfection method of AAV 
            vector production. It supplies the replication proteins from AAV2 
            and the engineered shh10 capsid protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8342)
  AUTHORS   Klimczak RR, Koerber JT, Dalkara D, Flannery JG, Schaffer DV
  TITLE     A novel adeno-associated viral variant for efficient and selective 
            intravitreal transduction of rat Muller cells.
  JOURNAL   PLoS One. 2009 Oct 14;4(10):e7467. doi: 
            10.1371/journal.pone.0007467.
  PUBMED    19826483
REFERENCE   2  (bases 1 to 8342)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-10-14"; volume: "4(10):e7467. doi"; pages: " 
            10.1371/journal.pone.0007467"
COMMENT     Sequence Label: shh10
FEATURES             Location/Qualifiers
     source          1..8342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(653..1166)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(848..869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1060..1079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2092..2103)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     repeat_region   6179..6281
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6353..6370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6524..7112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6604..6623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7283..8074,8075..8143))
                     /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     7906..7925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8144..8248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8316..8334
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcga
      241 ttccaacatc caataaatca tacaggcaag gcaaagaatt agcaaaatta agcaataaag
      301 cctcagagca taaagctaaa tcggttgtac caaaaacatt atgaccctgt aatacttttg
      361 cgggagaagc ctttatttca acgcaaggat aaaaattttt agaaccctca tatattttaa
      421 atgcaatgcc tgagtaatgt gtaggtaaag attcaaacgg gtgagaaagg ccggagacag
      481 tcaaatcacc atcaatatga tattcaaccg ttctagctga taaattcatg ccggagaggg
      541 tagctatttt tgagaggtct ctacaaaggc tatcaggtca ttgcctgaga gtctggagca
      601 aacaagagaa tcgatgaacg gtaatcgtaa aactagcatg tcaatcatat gtaccccggt
      661 tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt aaattgtaag
      721 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      781 atatgccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
      841 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      901 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
      961 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1021 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1081 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     1141 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgagcacg tataacgtgc
     1201 tttcctcgtt agaatcagag cgggagctaa acaggaggcc gattaaaggg attttagaca
     1261 ggaacggtac gccagaatcc tgagaagtgt ttttataatc agtgaggcca ccgagtaaaa
     1321 gagtctgtcc atcacgcaaa ttaaccgttg tcgcaatact tctttgatta gtaataacat
     1381 cacttgcctg agtagaagaa ctcaaactat cggccttgct ggtaatatcc agaacaatat
     1441 taccgccagc cattgcaacg gaatcgccat tcgccattca ggctgcgcaa ctgttgggaa
     1501 gggcgatcgg tgcgggcctc ttcgctatta cgccagcgat gcagcttctc ggccacggtc
     1561 aggggtgcct gctcaatcag attcagatcc atgtcagaat ctggcggcaa ctcccattcc
     1621 ttctcggcca cccagttcac aaagctgtca gaaatgccgg gcagatgctc gtcaaggtcg
     1681 ctggggacct taatcacaat ctcgtaaaac cccgccgtgg tggctgcgcg ttcaaacctc
     1741 ccgcttcaaa atggagaccc tgcgtgctca ctcgggctta aatacccagc gtgaccacat
     1801 ggtgtcgcaa aatgtcgcaa aacactcacg tgacctctaa tacaggacct ctagagcatg
     1861 gaaactagat aagaaagaaa tacgcagaga ccaaagttca actgaaacga attaaacggt
     1921 ttattgatta acaagcggcc gcaattacag gggacgggtg aggtaacggg tgccaatggg
     1981 gcgaggctca gtataaagtc cattgttgtc cacagtgaaa tcaacgttgg cagattttgc
     2041 atagttagat gtatactgca cttcgggatt ccagcgtttg ctgttttctt tctgcagctc
     2101 ccattcaatc tccacgctca cttgtcctgt ggaatactgg gtgatgaatg aagcaaactt
     2161 tgtagccgaa aactctgccg gaggattcgc aggaacaggc gtgtttttga tgaggatctg
     2221 aggaggcggg tgcttaagtc caaagccgcc catgagagga gacgggtgaa agtgtccatc
     2281 cgtgtgagga attttggccc aaataggacc ctgcaggtat acgtctctgt cttgccacac
     2341 cattccaggt aaggctccca taacatgcac atctccggtc gcagggtctg tgctgctgct
     2401 ctggagattg actgccacag tcccaaatct ttcggtggcc acggggttag tggctttgat
     2461 ttcctcttcg tctgtgatca tgacattgtc caatgcagtg tttgaagctc cggcgctctc
     2521 ctttccaaaa atcatgacac cgctcatggg aaagaacttg tttttgtcgt ctttgtgtga
     2581 ggccatagca gtgccagggt tgattataga ttcacgccca ttaaggttat attttgaagc
     2641 accagtccag gtaaagttgc tgttgttgtt gtctgttttt gttttagaaa cgcgctgctg
     2701 ccggtaacag ggtccaggta gccagttttt gggctgaaca gacatgccag ctggagaccc
     2761 ccggctaaac agcaagtcct tgttttgggc acttccggac tgatcctgag ttctgttcag
     2821 gtaatacagg tactggtcga tgagaggatt catcagccgg tccaggctct ggctgtgcgc
     2881 gtagctgctg tggaaaggca cgtcctcgaa ggtgtagctg aaggtaaagt tattgcccgt
     2941 tctcagcatc tgcgatggga aatattccag gcagtaaaag gatgaccgtc ccactgcctg
     3001 gctgccattg ttgagcgtta ggtagccgta ctgcggaatc atgaacacgt ccgccgggaa
     3061 cggagggagg cagccctggt gcgcagagcc gaggacgtac ggcaactggt actccgagtc
     3121 cgagaagact tgaaccgtgc tggtaaggtt attagcgatg gtcgtgacgc catcattcgt
     3181 cgtgacctcc ttgacttgga cgttgaagag cttgaagttg agtctcttgg gccggaatcc
     3241 ccaattgttg ttgatgagtc gctgccagtc acgtggtgag aaatggcagt ggaatctgtt
     3301 gaaatcaaaa tacccccagg gggtgctgta gccgaagtag tggttgtcgt tgctggcccc
     3361 cgttgaagca ctggagattt gcttgtagag gtggttgtta taggtgggca aggcccatgt
     3421 tcgggtgctg gtggtgatga ctctgtcgcc cagccatgtg gaatcgcaat gccaatttcc
     3481 tgaggcatta cccactccgt cggcgccttc gttattgtct gccattggtg cgccaccgcc
     3541 tgaagccatt gtagtaggtc ccacagcagc gggggttgct ggaggttctc cgagaggttg
     3601 tgggtcgggg actgactctg agtcgccagt ctgaccaaaa ttgagtctct ttttagcggg
     3661 ctgctggcct gtcttgccaa tgcccgagga ggagtctggc tcttgtggcg actgctctac
     3721 cggacgtttc tttccaggag ccgtcttagc accttcctca accagaccaa aaggttcgag
     3781 aaccctcttc ttggcctgga agactgctcg cccgaggttg cccccaaaag acgtatcttc
     3841 ttgcagacgc tcctgaaact cggcgtcggc gtggttatac cgcaggtacg gattgtcacc
     3901 cgctttgagc tgctggtcgt aggccttgtc gtgctcgagg gccgctgcat ccgccgcgtt
     3961 gacgggctcc cccttgtcga gtccgttgaa gggtccgagg tacttgtagc caggaagcac
     4021 cagaccccgg ccgtcgtcct gcttttgctg gttggctttg ggtttcgggg ctccaggttt
     4081 caagtcccac cactcgcgaa tgccctcaga gaggttgtcc tcaagccaat ctggaagata
     4141 accatcggca gccatacctg atttaaatca tttattgttc aaagatgcag tcatccaaat
     4201 ccacattgac cagatcgcag gcagtgcaag cgtctggcac ctttcccatg atatgatgaa
     4261 tgtagcacag tttctgatac gcctttttga cgacagaaac gggttgagat tctgacacgg
     4321 gaaagcactc taaacagtct ttctgtccgt gagtgaagca gatatttgaa ttctgattca
     4381 ttctctcgca ttgtctgcag ggaaacagca tcagattcat gcccacgtga cgagaacatt
     4441 tgttttggta cctgtctgcg tagttgatcg aagcttccgc gtctgacgtc gatggctgcg
     4501 caactgactc gcgcacccgt ttgggctcac ttatatctgc gtcactgggg gcgggtcttt
     4561 tcttggctcc accctttttg acgtagaatt catgctccac ctcaaccacg tgatcctttg
     4621 cccaccggaa aaagtctttg acttcctgct tggtgacctt cccaaagtca tgatccagac
     4681 ggcgggtgag ttcaaatttg aacatccggt cttgcaacgg ctgctggtgt tcgaaggtcg
     4741 ttgagttccc gtcaatcacg gcgcacatgt tggtgttgga ggtgacgatc acgggagtcg
     4801 ggtctatctg ggccgaggac ttgcatttct ggtccacgcg caccttgctt cctccgagaa
     4861 tggctttggc cgactccacg accttggcgg tcatcttccc ctcctcccac cagatcacca
     4921 tcttgtcgac acagtcgttg aagggaaagt tctcattggt ccagtttacg cacccgtaga
     4981 agggcacagt gtgggctatg gcctccgcga tgttggtctt cccggtagtt gcaggcccaa
     5041 acagccagat ggtgttcctc ttgccgaact ttttcgtggc ccatcccaga aagacggaag
     5101 ccgcatattg gggatcgtac ccgtttagtt ccaaaatttt ataaatccga ttgctggaaa
     5161 tgtcctccac gggctgctgg cccaccaggt agtcgggggc ggttttagtc aggctcataa
     5221 tctttcccgc attgtccaag gcagccttga tttgggaccg cgagttggag gccgcattga
     5281 aggagatgta tgaggcctgg tcctcctgga tccactgctt ctccgaggta atccccttgt
     5341 ccacgagcca cccgaccagc tccatgtacc tggctgaagt ttttgatctg atcaccggcg
     5401 catcagaatt gggattctga ttctctttgt tctgctcctg cgtctgcgac acgtgcgtca
     5461 gatgctgcgc caccaaccgt ttacgctccg tgagattcaa acaggcgctt aaatactgtt
     5521 ccatattagt ccacgcccac tggagctcag gctgggtttt ggggagcaag taattgggga
     5581 tgtagcactc atccaccacc ttgttcccgc ctccggcgcc atttctggtc tttgtgaccg
     5641 cgaaccagtt tggcaaagtc ggctcgatcc cgcggtaaat tctctgaatc agtttttcgc
     5701 gaatctgact caggaaacgt cccaaaacca tggatttcac cccggtggtt tccacgagca
     5761 cgtgcatgtg gaagtagctc tctcccttct caaattgcac aaagaaaagg gcctccgggg
     5821 ccttactcac acggcgccat tccgtcagaa agtcgcgctg cagcttctcg gccacggtca
     5881 ggggtgcctg ctcaatcaga ttcagatcca tgtcagaatc tggcggcaac tcccattcct
     5941 tctcggccac ccagttcaca aagctgtcag aaatgccggg cagatgctcg tcaaggtcgc
     6001 tggggacctt aatcacaatc tcgtaaaacc ccgccgtggt ggctgcgcgt tcaaacctcc
     6061 cgcttcaaaa tggagaccct gcgtgctcac tcgggcttaa atacccagcg tgaccacatg
     6121 gtgtcgcaaa atgtcgcaaa acactcacgt gacctctaat acaggacctc tagagctact
     6181 acgtcacccg ccccgttccc acgccccgcg ccacgtcaca aactccaccc cctcattatc
     6241 atattggctt caatccaaaa taaggtatat tattgatgat gggaattctg cattaatgaa
     6301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8281 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8341 gt
//
