LOCUS       sequence-448444-        8779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minigene encoding the CEP290 c.2991+1655A>G mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8779)
  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 8779)
  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.CEP290 c.2991+1655A>G
FEATURES             Location/Qualifiers
     source          1..8779
                     /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"
     regulatory      1779..1788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1788..1853
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(6483..6507)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6536..6657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6547..6566)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6601..6620
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6773..6790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6944..7532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7024..7043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7703..8494,8495..8563))
                     /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     8326..8345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8564..8668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8736..8754
                     /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 gggggggggg gggggsgggg sgrggcggrg 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 ckcgggccgg 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 caccatggac tacaaggacc
     1801 acgacggcga ctacaaagat catgatatcg attataagga cgatgacgat aaggaaatgg
     1861 ccattttcaa gattgcagct ctccaaaaag ttgtagataa tagtgtttct ttgtctgaac
     1921 tagaactggc taataaacag tacaatgaac tgactgctaa gtacagggac atcttgcaaa
     1981 aagataatat gcttgttcaa agaacaagta acttggaaca cctggaggta agtttgtgtg
     2041 attcttgaac cttgtgaaat tagccatttt tcttcaatat ttttgtgttt ggggggattt
     2101 ggcagatttt aattaaagtt tgcctgcatt tatataaatt taacagagat ataattatcc
     2161 atattattca ttcagtttag ttataaatat tttgttccca cataacacac acacacacac
     2221 acaatatatt atctatttat agtggctgaa tgacttctga atgattatct agatcattct
     2281 ccttaggtca cttgcatgat ttagctgaat caaacctctt ttaaccagac atctaagaga
     2341 aaaaggagca tgaaacaggt agaatattgt aatcaaagga gggaagcact cattaagtgc
     2401 ccatcccttt ctcttacccc tgtacccaga acaaactatt ctcccatggt ccctggcttt
     2461 tgttccttgg aatggatgta gccaacagta gctgaaatat taagggctct tcctggacca
     2521 tggatgcact ctgtaaattc tcatcatttt ttattgtaga ataaatgtag aattttaatg
     2581 tagaataaat ttatttaatg tagaataaaa aataaaaaaa ctagagtaga atatcataag
     2641 ttacaatctg tgaatatgga ccagaccctt tgtagttatc ttacagccac ttgaactcta
     2701 taccttttac tgaggacaga acaagctcct gatttgttca tcttcctcat cagaaataga
     2761 ggcttatgga ttttggatta ttcttatcta agatcctttc acaggagtag aataagatct
     2821 aattctatta gctcaaaagc ttttgctggc tcatagagac acattcagta aatgaaaacg
     2881 ttgttctgag tagctttcag gattcctact aaattatgag tcatgtttat caatattatt
     2941 tagaagtaat cataatcagt ttgctttctg ctgcttttgc caaagagagg tgattatgtt
     3001 actttttata gaaaattatg cctatttagt gtggtgataa tttatttttt tccattctcc
     3061 atgtcctctg tcctatcctc tccagcatta gaaagtccta ggcaagagac atcttgtgga
     3121 taatgtatca atgagtgatg tttaacgtta tcattttccc aaagagtatt tttcatcttt
     3181 cctaaagatt tttttttttt ttttttgaga tggagtttca ttctgtcacc caggctgagt
     3241 gcagtggcac gatctcggct taacgcttac tgcatcctct gcctcccaga ttcaagcagt
     3301 tctcctgcct cagcctctga gtagctggga ttacaggtgt gcaccaccac accagctaat
     3361 tttttttttt tttttttttt ttttgaggca gagtctcgct ctgtcaccca ggctggagtg
     3421 cagtggcgcc atcttggctc actgcaagct ccacctcccg ggttcaggcc gttctcctgc
     3481 ctcagcctcc tgagtagctg gtaccacagg cacccaccat catgcccggc taattttttg
     3541 tatttttagt agagatgggg tttcaccttg ttagccagga tggtgtcgat ctcctgaact
     3601 cgtgatccac ccgcctcggc ctcctaaagt gctgggatta cagatgtgag ccaccgcacc
     3661 tggccccagt tgtaattgtg agtatctcat acctatccct attggcagtg tcttagtttt
     3721 attttttatt atctttattg tggcagccat tattcctgtc tctatctcca gtcttacatc
     3781 ctccttactg ccacaagaat gatcattcta aacatgaatc ctaccctgtg actcccatgt
     3841 gactccccgc cttaaaaact gtcaaaagct accggttacc tgaagggtaa aagtcaagtc
     3901 ccctacttac ctcatgtcat ctagagcaag agatgaacta gctgagtttt ctgaccacag
     3961 tgttctttct tatgtatgtt cttttgtacg tgctcttttc tatatatagg gaaccatttc
     4021 tctcttccag ttgttttgct cagtgaattt ctattcctgt ttcaaaactt gttcaggcat
     4081 tacctttttt ttcttaagca tacttttttt aatggaacaa agtcactcct gtctacacta
     4141 gttctgcatc ttatacatag gttttgtaca tagtacatat ttatatcaca tcaaattata
     4201 tgtgtttaca tatctgtctt ccttaatgga atataagtct tttgatataa ggaactattt
     4261 aatttgtttc tgtgtgttga gtatctcctg tttggcacag agttcaagct aatacatgag
     4321 agtgattagt ggtggagagc cacagtgcat gtggtgtcaa atatggtgct taggaaatta
     4381 ttgttgcttt ttgagaggta aaggttcatg agactagagg tcacgaaaat cagatttcat
     4441 gtgtgaagaa tggaatagat aataaggaaa tacaaaaact ggatgggtaa taaagcaaaa
     4501 gaaaaacttg aaatttgata gtagaagaaa aaagaaatag atgtagattg aggtagaatc
     4561 aagaagagga ttcttttttt gttgtttttt tttttgaaac agagtctcac tgtgttgccc
     4621 aggctggagt gcagtggagt gatcttggct tactgcaacc tctgcctccc aggttcaagc
     4681 gattcttctg cttcagtctc ccgagtagct ggaattacag gtgcccacca gcacggccgg
     4741 ctaatttagt agagacaggg ttttgccatg ttggccgggc tggtctcaaa ctttggatct
     4801 caggtaatcc gccagcctca acttcccaaa gtgctgggat tacaggcatg agccactgtg
     4861 cccagcctgt tttttttttt ttaaaggaga ccagtgaagt ttcaggagga gggaaagaaa
     4921 atttagagtt actagggaga gagtgatgaa gataagagat gaaagtggta ataagggaaa
     4981 tagcaaaata tcagggtagg tgggagaaaa agagatttgt aacaaacaat aggattatcc
     5041 tgtgaaaaag gatgaaagga agaaaaaaat ggatagaaag atatttaaaa caccctcagc
     5101 ctcctgtttt ccctcctgtg tattcatagt atataaaact ataattatgt actttactta
     5161 aaaaatatat tattattacc ttatcgtgct tatttaatca tagcatgtcc tctttttagt
     5221 ctcattaccc tgtttgtatt attcttcata acacttaata cctgacattg tattatatat
     5281 tggcttattt tccaggtact ccactcaaat ataagttcta ggatataatt tatttatcac
     5341 tgaaatccat tgcttagagt acctggcatg tagtaaatag gcattctgtt ttttcaaata
     5401 aaaaataaag gaacttaaga tatatattta tgttatatcg ccagcctttt tcctcacagc
     5461 tctattctgt tgtacagaat tacctacttt acaattcctg tgtttcaagg ggatctcaaa
     5521 tttaacgtgt ccacaatgaa ctcctgattt ctgtttctct cctagtcatt cttatttcaa
     5581 tatatgttca gttacctaac cagctagtca aggcagatac tttagagtta ttctgtagtc
     5641 attctttttc cctaccattt ttgttttcca aatgtaattt atgtgtgtct tcttcatcct
     5701 cgcagctcta acccttgtcc aaaccagcat catcactcat ctggagttcc acaatgtctt
     5761 tctggctagt ttccctgatt tctctattga cccctttatt ctccacagtg cagccagaat
     5821 gattgtttaa aacttcctcc ttaaaatctt taaattgttt tcttttatac gttaagttaa
     5881 attccagttc cttgtcttgg catgccatgc cctgcctggt gtggcccctg atggtctctc
     5941 caacttcatg ttttactact attgactctt atttttgctt actctgcttg ggtgctccag
     6001 tcctccaaat catttcctgc tccaatcatt tcaatcattt tttcctctca gatcttatag
     6061 tattccaaat gctttcttcc tttggagcat ctgggtttac taataaatac ttcgtacctc
     6121 acagttcagc ttaaatatca attatttggt ggttaagaca tccttcaacc gctctatcta
     6181 aatgttcctt tctattattc actggctcag tactctgttt ttattttctt tctaaatgtc
     6241 aacttttttt tttttgagtc agggtctcac tgttgcccag gctcgagtgc agttgcacaa
     6301 tcatagctca ttgcattggt ctgaattaca atgtagtttc ttagtgatat ttttccttta
     6361 ttcagtgtga aaacatctcc ttaaaagaac aagtggagtc tataaataaa gaactggaga
     6421 ttaccaagga aaaacttcac actattgaac aagcctggga acaggaaact aaattagatt
     6481 aaacccagct ttcttgtaca aagtggtgat ggccggccgc ttcgagcaga catgataaga
     6541 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6601 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     6661 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     6721 agcaagtaaa acctctacaa atgtggtagc ggccgcggcg ctcttccgct tcctcgctca
     6781 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6841 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6901 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6961 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7021 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7081 tgccgcttac cggatacctg tccgcctttc tctcttcggg aagcgtggcg ctttctcata
     7141 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7201 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7261 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7321 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7381 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7441 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7501 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7561 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7621 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7681 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7741 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7801 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7861 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7921 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7981 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8041 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8101 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8161 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8221 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8281 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8341 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8401 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8461 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8521 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8581 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8641 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8701 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8761 gtcggcgcgc cgcggccgc
//
