LOCUS       sequence-342083-        8369 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 7M8 capsid protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8369)
  AUTHORS   Dalkara D, Byrne LC, Klimczak RR, Visel M, Yin L, Merigan WH, 
            Flannery JG, Schaffer DV
  TITLE     In vivo-directed evolution of a new adeno-associated virus for 
            therapeutic outer retinal gene delivery from the vitreous.
  JOURNAL   Sci Transl Med. 2013 Jun 12;5(189):189ra76. doi: 
            10.1126/scitranslmed.3005708.
  PUBMED    23761039
REFERENCE   2  (bases 1 to 8369)
  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: "Sci Transl 
            Med."; date: "2013-06-12"; volume: "5(189):189ra76. doi"; pages: " 
            10.1126/scitranslmed.3005708"
COMMENT     Sequence Label: 7M8
FEATURES             Location/Qualifiers
     source          1..8369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   8..110
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     CDS             4213..4224
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     rep_origin      5150..5663
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5237..5256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5447..5468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6145..6164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6264..6286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6324..6342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6410..6514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6515..6583,6584..7375)
                     /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(6733..6752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7546..8134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8035..8054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8288..8305
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaattcccat catcaataat ataccttatt ttggattgaa gccaatatga taatgagggg
       61 gtggagtttg tgacgtggcg cggggcgtgg gaacggggcg ggtgacgtag tagctctaga
      121 ggtcctgtat tagaggtcac gtgagtgttt tgcgacattt tgcgacacca tgtggtcacg
      181 ctgggtattt aagcccgagt gagcacgcag ggtctccatt ttgaagcggg aggtttgaac
      241 gcgcagccac cacggcgggg ttttacgaga ttgtgattaa ggtccccagc gaccttgacg
      301 agcatctgcc cggcatttct gacagctttg tgaactgggt ggccgagaag gaatgggagt
      361 tgccgccaga ttctgacatg gatctgaatc tgattgagca ggcacccctg accgtggccg
      421 agaagctgca gcgcgacttt ctgacggaat ggcgccgtgt gagtaaggcc ccggaggccc
      481 ttttctttgt gcaatttgag aagggagaga gctacttcca catgcacgtg ctcgtggaaa
      541 ccaccggggt gaaatccatg gttttgggac gtttcctgag tcagattcgc gaaaaactga
      601 ttcagagaat ttaccgcggg atcgagccga ctttgccaaa ctggttcgcg gtcacaaaga
      661 ccagaaatgg cgccggaggc gggaacaagg tggtggatga gtgctacatc cccaattact
      721 tgctccccaa aacccagcct gagctccagt gggcgtggac taatatggaa cagtatttaa
      781 gcgcctgttt gaatctcacg gagcgtaaac ggttggtggc gcagcatctg acgcacgtgt
      841 cgcagacgca ggagcagaac aaagagaatc agaatcccaa ttctgatgcg ccggtgatca
      901 gatcaaaaac ttcagccagg tacatggagc tggtcgggtg gctcgtggac aaggggatta
      961 cctcggagaa gcagtggatc caggaggacc aggcctcata catctccttc aatgcggcct
     1021 ccaactcgcg gtcccaaatc aaggctgcct tggacaatgc gggaaagatt atgagcctga
     1081 ctaaaaccgc ccccgactac ctggtgggcc agcagcccgt ggaggacatt tccagcaatc
     1141 ggatttataa aattttggaa ctaaacgggt acgatcccca atatgcggct tccgtctttc
     1201 tgggatgggc cacgaaaaag ttcggcaaga ggaacaccat ctggctgttt gggcctgcaa
     1261 ctaccgggaa gaccaacatc gcggaggcca tagcccacac tgtgcccttc tacgggtgcg
     1321 taaactggac caatgagaac tttcccttca acgactgtgt cgacaagatg gtgatctggt
     1381 gggaggaggg gaagatgacc gccaaggtcg tggagtcggc caaagccatt ctcggaggaa
     1441 gcaaggtgcg cgtggaccag aaatgcaagt cctcggccca gatagacccg actcccgtga
     1501 tcgtcacctc caacaccaac atgtgcgccg tgattgacgg gaactcaacg accttcgaac
     1561 accagcagcc gttgcaagac cggatgttca aatttgaact cacccgccgt ctggatcatg
     1621 actttgggaa ggtcaccaag caggaagtca aagacttttt ccggtgggca aaggatcacg
     1681 tggttgaggt ggagcatgaa ttctacgtca aaaagggtgg agccaagaaa agacccgccc
     1741 ccagtgacgc agatataagt gagcccaaac gggtgcgcga gtcagttgcg cagccatcga
     1801 cgtcagacgc ggaagcttcg atcaactacg cagacaggta ccaaaacaaa tgttctcgtc
     1861 acgtgggcat gaatctgatg ctgtttccct gcagacaatg cgtgagaatg aatcagaatt
     1921 caaatatctg cttcactcac ggacagaaag actgtttaga gtgctttccc gtgtcagaat
     1981 ctcaacccgt ttctgtcgtc aaaaaggcgt atcagaaact gtgctacatt catcatatca
     2041 tgggaaaggt gccagacgct tgcactgcct gcgatctggt caatgtggat ttggatgact
     2101 gcatctttga acaataaatg atttaaatca ggtatggctg ccgatggtta tcttccagat
     2161 tggctcgagg acactctctc tgaaggaata agacagtggt ggaagctcaa acctggccca
     2221 ccaccaccaa agcccgcaga gcggcataag gacgacagca ggggtcttgt gcttcctggg
     2281 tacaagtacc tcggaccctt caacggactc gacaagggag agccggtcaa cgaggcagac
     2341 gccgcggccc tcgagcacga caaagcctat gaccggcagc tcgacagcgg agacaacccg
     2401 tacctcaagt acaaccacgc cgacgcggag tttcaggagc gccttaaaga agatacgtct
     2461 tttgggggca acctcggacg agcagtcttc caggcgaaaa agagggttct tgaacctctg
     2521 ggcctggttg aggaacctgt taagacggct ccgggaaaaa agaggccggt agagcactct
     2581 cctgtggagc cagactcctc ctcgggaacc ggaaaggcgg gccagcagcc tgcaagaaaa
     2641 agattgaatt ttggtcagac tggagacgca gactcagtac ctgaccccca gcctctcgga
     2701 cagccaccag cagccccctc tggtctggga actaatacga tggctacagg cagtggcgca
     2761 ccaatggcag acaataacga gggcgccgac ggagtgggta attcctcggg aaattggcat
     2821 tgcgattcca catggatggg cgacagagtc accaccacca gcacccgaac ctgggccctg
     2881 cccacctaca acaaccacct ctacaaacaa atttccagcc aatcaggagc ctcgaacgac
     2941 aatcactact ttggctacag caccccttgg gggtattttg acttcaacag attccactgc
     3001 cacttttcac cacgtgactg gcaaagactc atcaacaaca actggggatt ccgacccaag
     3061 agactcaact tcaagctctt taacattcaa gtcaaagagg tcacgcagaa tgacggtacg
     3121 acgacgattg ccaataacct taccagcacg gttcaggtgt ttactgactc ggagtaccag
     3181 ctcccgtacg tcctcggctc ggcgcatcaa ggatgcctcc cgccgttccc agcagacgtc
     3241 ttcatggtgc cacagtatgg atacctcacc ctgaacaacg ggagtcaggc agtaggacgc
     3301 tcttcatttt actgcctgga gtactttcct tctcagatgc tgcgtaccgg aaacaacttt
     3361 accttcagct acacttttga ggacgttcct ttccacagca gctacgctca cagccagagt
     3421 ctggaccgtc tcatgaatcc tctcatcgac cagtacctgt attacttgag cagaacaaac
     3481 actccaagtg gaaccaccac gcagtcaagg cttcagtttt ctcaggccgg agcgagtgac
     3541 attcgggacc agtctaggaa ctggcttcct ggaccctgtt accgccagca gcgagtatca
     3601 aagacatctg cggataacaa caacagtgaa tactcgtgga ctggagctac caagtaccac
     3661 ctcaatggca gagactctct ggtgaatccg ggcccggcca tggcaagcca caaggacgat
     3721 gaagaaaagt tttttcctca gagcggggtt ctcatctttg ggaagcaagg ctcagagaaa
     3781 acaaatgtgg acattgaaaa ggtcatgatt acagacgaag aggaaatcag gacaaccaat
     3841 cccgtggcta cggagcagta tggttctgta tctaccaacc tccagagagg caacctagca
     3901 ctcggcgaaa caacaagacc tgctaggcaa gcagctaccg cagatgtcaa cacacaaggc
     3961 gttcttccag gcatggtctg gcaggacaga gatgtgtacc ttcaggggcc catctgggca
     4021 aagattccac acacggacgg acattttcac ccctctcccc tcatgggtgg attcggactt
     4081 aaacaccctc ctccccagat tctcatcaag aacaccccgg tacctgcgaa tccttcgacc
     4141 accttcagtg cggcaaagtt tgcttccttc atcacacagt actccacggg acaggtcagc
     4201 gtggagatcg agtgggagct gcagaaggaa aacagcaaac gctggaatcc cgaaattcag
     4261 tacacttcca actacaacaa gtctattaat gtggacttta ctgtggacac taatggcgtg
     4321 tattcagagc ctcgccccat tggcaccaga tacctgactc gtaatctgta attgcggccg
     4381 cttgttaatc aataaaccgt ttaattcgtt tcagttgaac tttggtctct gcgtatttct
     4441 ttcttatcta gtttccatgc tctagaggtc ctgtattaga ggtcacgtga gtgttttgcg
     4501 acattttgcg acaccatgtg gtcacgctgg gtatttaagc ccgagtgagc acgcagggtc
     4561 tccattttga agcgggaggt ttgaacgcgc agccaccacg gcggggtttt acgagattgt
     4621 gattaaggtc cccagcgacc ttgacgagca tctgcccggc atttctgaca gctttgtgaa
     4681 ctgggtggcc gagaaggaat gggagttgcc gccagattct gacatggatc tgaatctgat
     4741 tgagcaggca cccctgaccg tggccgagaa gctgcatcgc tggcgtaata gcgaagaggc
     4801 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gattccgttg
     4861 caatggctgg cggtaatatt gttctggata ttaccagcaa ggccgatagt ttgagttctt
     4921 ctactcaggc aagtgatgtt attactaatc aaagaagtat tgcgacaacg gttaatttgc
     4981 gtgatggaca gactctttta ctcggtggcc tcactgatta taaaaacact tctcaggatt
     5041 ctggcgtacc gttcctgtct aaaatccctt taatcggcct cctgtttagc tcccgctctg
     5101 attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc aaccatagta cgcgccctgt
     5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5221 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5341 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5461 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5521 ccgatttcgg catattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5581 aacaaaatat taacgcttac aatttaaata tttgcttata caatcttcct gtttttgggg
     5641 cttttctgat tatcaaccgg ggtacatatg attgacatgc tagttttacg attaccgttc
     5701 atcgattctc ttgtttgctc cagactctca ggcaatgacc tgatagcctt tgtagagacc
     5761 tctcaaaaat agctaccctc tccggcatga atttatcagc tagaacggtt gaatatcata
     5821 ttgatggtga tttgactgtc tccggccttt ctcacccgtt tgaatcttta cctacacatt
     5881 actcaggcat tgcatttaaa atatatgagg gttctaaaaa tttttatcct tgcgttgaaa
     5941 taaaggcttc tcccgcaaaa gtattacagg gtcataatgt ttttggtaca accgatttag
     6001 ctttatgctc tgaggcttta ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt
     6061 tattggatgt tggaatcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6121 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6181 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6241 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6301 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6361 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6481 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6541 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6601 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6661 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6721 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6781 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6841 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6901 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6961 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7021 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7081 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7141 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7201 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7261 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7321 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7381 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8221 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     8281 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     8341 ccccgcgcgt tggccgattc attaatgca
//
