LOCUS       sequence-448442-        6210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minigene encoding USH2A exon 13 and surrounding introns/exons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6210)
  AUTHORS   Tabaglio T, Agarwal T, Cher WY, Ow JR, Chew AK, Sun PYQ, Reddy 
            Gurrampati RS, Lu H, Naidu P, Ng HK, Le Guezennec X, Ng SY, 
            Lakshmanan M, Guccione E, Wee KB
  TITLE     Unveiling sequence-agnostic mixed-chemical modification patterns for
            splice-switching oligonucleotides using the NATURA platform.
  JOURNAL   Mol Ther Nucleic Acids. 2025 Jan 7;36(1):102422. doi: 
            10.1016/j.omtn.2024.102422. eCollection 2025 Mar 11.
  PUBMED    39926316
REFERENCE   2  (bases 1 to 6210)
  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: "Mol Ther 
            Nucleic Acids."; date: "2025-01-7"; volume: "36(1):102422. doi"; 
            pages: " 10.1016/j.omtn.2024.102422. eCollection 2025 Mar 11"
COMMENT     Sequence Label: pRP.USH2A
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        25..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          684..1700
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1624..1646
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1708..1727
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1754..1778
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3914..3938)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3967..4088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3978..3997)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4032..4051
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4204..4221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4375..4963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4455..4474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5134..5925,5926..5994))
                     /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     5757..5776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5995..6099)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6167..6185
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt gctcgacatt gattattgac tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      421 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      481 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccggggg
      541 gggggggggg gggsgggggg gggggsgggg sgrggsggrg aggkgcggcg gcagccaatc
      601 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      661 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      721 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      781 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      841 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      901 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
      961 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1021 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1081 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     1141 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     1201 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     1261 aggkgggggt gccgggcggg gcggggccgc cycgggccgg ggrgggctcg ggggrggggc
     1321 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     1381 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     1441 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     1501 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     1561 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     1621 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     1681 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1741 ttggcaaaga attgcaagtt tgtacaaaaa agcaggctgc caccatgatt ggaggacagt
     1801 gtaattgtaa gagacacgtg tctggcaggc agtgcaatca gtgccagaat ggattctaca
     1861 atctacaaga gttggatcct gatggctgca gtccctgtaa ctgcaatacc tctgggacag
     1921 tggatggaga tattacctgt caccaaaatt caggccagtg caagtgcaaa gcaaacgtta
     1981 ttggtacgtg taacgcaaaa gtttacagtc acatctctat taccatctgg aatttattat
     2041 taagtatagt caccatgctg tacaatagag ctccagaact tattcattct gtctagctga
     2101 aactttgtac tcagcttaac cttttattaa acatctttag agatttctta tctttagaaa
     2161 aacaactaat ttgttatatg taattctact ataattttaa atgagcacat ttgttaaaat
     2221 agtttttaag atttgttaaa gagaaaaaga gctccagcat atgtaacaga aacaacattt
     2281 gcattaagca tttttctttg cattaagtaa taattaaaaa tttatgaagt tcatcgcaaa
     2341 cagttgtata ttaaagctaa attaaatatt gtcattgaat tttgagagta agattggccc
     2401 cctatggcat tgcttgtgag aaaacactca atattttgtg ttcgtatcat ctgcagtagc
     2461 attgtttgtg tctcgtctat cttgaatgaa atcattttcc catcctcacc ttttaaatat
     2521 attttatctt tagggcttag gtgtgatcat tgcaattttg gatttaaatt tctccgaagc
     2581 tttaatgatg ttggatgtga gccctgccag tgtaacctcc atggctcagt gaacaaattc
     2641 tgcaatcctc actctgggca gtgtgagtgc aaaaaagaag ccaaaggact tcagtgtgac
     2701 acctgcagag aaaactttta tgggttagat gtcaccaatt gtaaggcctg tgactgtgac
     2761 acagctggat ccctccctgg gactgtctgt aatgctaaga cagggcagtg catctgcaag
     2821 cccaatgttg aagggagaca gtgcaataaa tgtttggagg gaaacttcta cctacggcaa
     2881 aataattctt tcctctgtct gccttgcaac tgtgataaga ctgggacaat aaatggctct
     2941 ctgctgtgta acaaatcaac aggacaatgt ccttgcaaat taggggtaac aggtcttcgc
     3001 tgtaatcagt gtgagcctca caggtacaat ttgaccattg acaattttca acactgccag
     3061 atgtgtgagt gtgattcctt ggggacatta cctgggacca tttgtgaccc aatcagtggc
     3121 cagtgcctgt gtgtgcctaa tcgtcaagga agaaggtgta atcagtgtca accaggtaag
     3181 aaagaaatgt attacatttt cagtgcacaa tgacattcct tttgttaact taggtaactt
     3241 ctccctgttt ctggtttgtg gcttctacaa attttatttc caaaatcatt actgtattta
     3301 tatcattatc caacacatat ataactattt aacttattca aaattatctg catatttatg
     3361 ttactatttt gagaggatac tttagataaa actcagccga tcggatttat ttcataattg
     3421 agactcaatt tctacacttg aagtaaatct cctttttaac agttttttaa aaatcagatc
     3481 aacaagagtc aattttattt tccagagaaa ggaaaatttg agttgaatat ccatacaatg
     3541 ccaaatattc aaatgatgaa ctaaatctct gaataaagct ggctaaatgt ttttgctgaa
     3601 gaggctatat gttctagttt tatatagaaa tacctagaat tgtttccaca tgccatcaaa
     3661 ttaataaaat aggccaaaga gtggataaaa agtatatata aaacttttaa tttctttatt
     3721 tctaggtttt tatatttctc caggcaatgc cactggctgc ctgccatgct catgccatac
     3781 aactggtgca gttaatcaca tctgtaatag cctgactggt cagtgtgttt gccaagatgc
     3841 ttccattgct gggcaacgtt gtgaccaatg caaagaccat tactttggat ttgatcctca
     3901 gactggaaga tagacccagc tttcttgtac aaagtggtga tggccggccg cttcgagcag
     3961 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4021 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4081 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4141 aggtttttta aagcaagtaa aacctctaca aatgtggtag cggccgcggc gctcttccgc
     4201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctctcttcgg gaagcgtggc
     4561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6121 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6181 gaggcccttt cgtcggcgcg ccgcggccgc
//
