LOCUS       sequence-372743-        6398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  minigene assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  AUTHORS   Pavinato L, Stanic J, Barzasi M, Gurgone A, Chiantia G, Cipriani V, 
            Eberini I, Palazzolo L, Di Luca M, Costa A, Marcantoni A, Biamino E,
            Spada M, Hiatt SM, Kelley WV, Vestito L, Sisodiya SM, Efthymiou S, 
            Chand P, Kaiyrzhanov R, Bruselles A, Cardaropoli S, Tartaglia M, De 
            Rubeis S, Buxbaum JD, Smedley D, Ferrero GB, Giustetto M, Gardoni F,
            Brusco A
  TITLE     Missense variants in RPH3A cause defects in excitatory synaptic 
            function and are associated with a clinically variable 
            neurodevelopmental disorder.
  JOURNAL   Genet Med. 2023 Nov;25(11):100922. doi: 10.1016/j.gim.2023.100922. 
            Epub 2023 Jul 1.
  PUBMED    37403762
REFERENCE   2  (bases 1 to 6398)
  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: "Genet 
            Med."; date: "2023-11"; volume: "25(11):100922. doi"; pages: " 
            10.1016/j.gim.2023.100922. Epub 2023 Jul 1"
COMMENT     Sequence Label: pSPL3-hRPH3A_c.1853Gmut
FEATURES             Location/Qualifiers
     source          1..6398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..25)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10..339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      190..325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    2853..3086
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3271..3315
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     polyA_signal    3630..3764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3654..3673)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3708..3727
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4438..4542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4543..4611,4612..5403)
                     /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(4761..4780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5574..6162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6063..6082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6316..6333
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
       61 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
      121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
      181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggact gtgtttactt
      361 gcaatccccc aaaacagaca gaatggtgca tctgtccagt gaggagaagt ctgcggtcac
      421 tgccctgtgg ggcaaggtga atgtggaaga agttggtggt gaggccctgg gcaggctgct
      481 ggttgtctac ccatggaccc agaggttctt cgagtccttt ggggacctgt cctctgcaaa
      541 tgctgttatg aacaatccta aggtgaaggc tcatggcaag aaggtgctgg ctgccttcag
      601 tgagggtctg agtcacctgg acaacctcaa aggcaccttt gctaagctga gtgaactgca
      661 ctgtgacaag ctgcacgctc tagagtcgac ccagcagtaa gtaatacatg taatgcaacc
      721 tatacaaata gcaatagtag cattagtagt agcaataata atagcaatag ttgtgtggtc
      781 catagtaatc atagaatata ggaaaatatt aagacaaaga aaaatagaca ggttaattga
      841 tagactaata gaaagagcag aagacagtgg caatgagagt gaaggagaaa tatcagcact
      901 tgtggagatg ggggtggaga tggggcacca tgctccttgg gatgttgatg atctgtagtg
      961 ctacagaaaa attgtgggtc acagtctatt atggggtagg gatcaccaga attctggagc
     1021 tcgaggatgc tctaatgcat attggtcatt atcaatgttg atccctttct gatttattca
     1081 tcaattcacc ctgataaaga aggggccttt ggggtcaaga ggctgttctt attcattaaa
     1141 cgtcatccat aatagcatgt tgtgtcgcct cccagctggc tgaaaccgga catgggaaag
     1201 aaggccaaac acaagactca aattaaaaag aaaaccttga atcccgaatt cagtgaggta
     1261 aggctgccct attctttttc atgctctggg atatttgctg tgtgttagag aggcacaaat
     1321 agccacatag ccattaaata tgtggccaca tagccattaa atgcagccat ttgaaatgat
     1381 gatgtagaaa aatatttgaa aaatattgga tcccagatat ctggtgatcc cgtacctgtg
     1441 tggaaggaag caaccaccac tctattttgt gcatcagatg ctaaagcata tgatacagag
     1501 gtacataatg tttgggccac acatgcctgt gtacccacag accccaaccc acaagaagta
     1561 gtattggtaa atgtgacaga aaattttaac atgtggaaaa atgacatggt agaacagatg
     1621 catgaggata taatcagttt atgggatcaa agcctaaagc catgtgtaaa attaacccca
     1681 ctctgtgtta gtttaaagtg cactgatttg aagaatgata ctaataccaa tagtagtagc
     1741 gggagaatga taatggagaa aggagagata aaaaactgct ctttcaatat cagcacaagc
     1801 ataagaggta aggtgcagaa agaatatgca tttttttata aacttgatat aataccaata
     1861 gataatgata ctaccagcta tacgttgaca agttgtaaca cctcagtcat tacacaggcc
     1921 tgtccaaagg tatcctttga gccaattccc atacattatt gtgccccggc tggttttgcg
     1981 attctaaaat gtaataataa gacgttcaat ggaacaggac catgtacaaa tgtcagcaca
     2041 gtacaatgta cacatggaat taggccagta gtatcaactc aactgctgtt aaatggcagt
     2101 ctagcagaag aagaggtagt aattagatct gtcaatttca cggacaatgc taaaaccata
     2161 atagtacagc tgaacacatc tgtagaaatt aattgtacaa gacccaacaa caatacaaga
     2221 aaaaaaatcc gtatccagag gggaccaggg agagcatttg ttacaatagg aaaaatagga
     2281 aatatgagac aagcacattg taacattagt agagcaaaat ggaatgccac tttaaaacag
     2341 atagctagca aattaagaga acaatttgga aataataaaa caataatctt taagcaatcc
     2401 tcaggagggg acccagaaat tgtaacgcac agttttaatt gtggagggga atttttctac
     2461 tgtaattcaa cacaactgtt taatagtact tggtttaata gtacttggag tactgaaggg
     2521 tcaaataaca ctgaaggaag tgacacaatc acactcccat gcagaataaa acaatttata
     2581 aacatgtggc aggaagtagg aaaagcaatg tatgcccctc ccatcagcgg acaaattaga
     2641 tgttcatcaa atattacagg gctgctatta acaagagatg gtggtaataa caacaatggg
     2701 tccgagatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     2761 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     2821 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     2881 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     2941 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     3001 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     3061 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     3121 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     3181 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     3241 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     3301 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     3361 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctgta
     3421 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacctggag atctcccgag
     3481 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     3541 cattcgacca attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     3601 ccaatgccct ggctcacaaa taccactgag atccagacat gataagatac attgatgagt
     3661 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3721 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3781 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3841 tctacaaatg tggtatggct gattatgatc cccaggaagc tcctctgtgt cctcataaac
     3901 cctaacctcc tctacttgag aggacattcc aatcataggc tgcccatcca ccctctgtgt
     3961 cctcctgtta attaggtcac ttaacaaaaa ggaaattggg taggggtttt tcacagaccg
     4021 ctttctaagg gtaattttaa aatatctggg aagtcccttc cactgctgtg ttccagaagt
     4081 gttggtaaac agcccacaaa tgtcaacagc agaaacatac aagctgtcag ctttgcacaa
     4141 gggcccaaca ccctgctcat caagaagcac tgtggttgct gtgttagtaa tgtgcaaaac
     4201 aggaggcaca ttttccccac ctgtgtaggt tccaaaatat ctagtgtttt catttttact
     4261 tggatcagga acccagcact ccactggata agcattatcc ttatccaaaa cagccttgtg
     4321 gtcagtgttc atctgctgac tgtcaactgt agcatttttt ggggttacag tttgagcagg
     4381 atatttggtc ctgtagtttg ctaacacacc ccaggtggca cttttcgggg aaatgtgcgc
     4441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6361 ccgcctctcc ccgcgcgttg gccgattcat taatgcag
//
