LOCUS       sequence-372349-        6400 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 6400)
  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 6400)
  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.444Tmut
FEATURES             Location/Qualifiers
     source          1..6400
                     /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    2855..3088
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3273..3317
                     /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    3632..3766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3656..3675)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3710..3729
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4440..4544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4545..4613,4614..5405)
                     /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(4763..4782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5576..6164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6065..6084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6318..6335
                     /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 tcgagattct aactggcctt gctcatagaa ggaagtggga gaaatggaag caagttcttc
     1081 ctctttacat ccaccaagag aaagtggggg gctacgttgc catgcctccc atgagcatgg
     1141 ctcatctgag tgtgttggct gtgtttcatc cacagaacgt ctgcaccaag tgcggagtgg
     1201 agaccaacaa ccgcctgcat tctgtgtggc tctgcaaaat ctgcattgag cagagggatg
     1261 tgagtgccct ggtcccacct ggtgcctaga tcaccctcct ttcttggcca gcttaagagg
     1321 tgccttaaga ggtttgtatg aaaggaccca gctcagcagt gaacattgac agaacaacct
     1381 cagagtattt ggatgaatta tttctcctcg gatcccagat atctggtgat cccgtacctg
     1441 tgtggaagga agcaaccacc actctatttt gtgcatcaga tgctaaagca tatgatacag
     1501 aggtacataa tgtttgggcc acacatgcct gtgtacccac agaccccaac ccacaagaag
     1561 tagtattggt aaatgtgaca gaaaatttta acatgtggaa aaatgacatg gtagaacaga
     1621 tgcatgagga tataatcagt ttatgggatc aaagcctaaa gccatgtgta aaattaaccc
     1681 cactctgtgt tagtttaaag tgcactgatt tgaagaatga tactaatacc aatagtagta
     1741 gcgggagaat gataatggag aaaggagaga taaaaaactg ctctttcaat atcagcacaa
     1801 gcataagagg taaggtgcag aaagaatatg cattttttta taaacttgat ataataccaa
     1861 tagataatga tactaccagc tatacgttga caagttgtaa cacctcagtc attacacagg
     1921 cctgtccaaa ggtatccttt gagccaattc ccatacatta ttgtgccccg gctggttttg
     1981 cgattctaaa atgtaataat aagacgttca atggaacagg accatgtaca aatgtcagca
     2041 cagtacaatg tacacatgga attaggccag tagtatcaac tcaactgctg ttaaatggca
     2101 gtctagcaga agaagaggta gtaattagat ctgtcaattt cacggacaat gctaaaacca
     2161 taatagtaca gctgaacaca tctgtagaaa ttaattgtac aagacccaac aacaatacaa
     2221 gaaaaaaaat ccgtatccag aggggaccag ggagagcatt tgttacaata ggaaaaatag
     2281 gaaatatgag acaagcacat tgtaacatta gtagagcaaa atggaatgcc actttaaaac
     2341 agatagctag caaattaaga gaacaatttg gaaataataa aacaataatc tttaagcaat
     2401 cctcaggagg ggacccagaa attgtaacgc acagttttaa ttgtggaggg gaatttttct
     2461 actgtaattc aacacaactg tttaatagta cttggtttaa tagtacttgg agtactgaag
     2521 ggtcaaataa cactgaagga agtgacacaa tcacactccc atgcagaata aaacaattta
     2581 taaacatgtg gcaggaagta ggaaaagcaa tgtatgcccc tcccatcagc ggacaaatta
     2641 gatgttcatc aaatattaca gggctgctat taacaagaga tggtggtaat aacaacaatg
     2701 ggtccgagat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     2761 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     2821 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     2881 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     2941 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     3001 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     3061 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     3121 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     3181 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     3241 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     3301 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     3361 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttctg
     3421 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacctgg agatctcccg
     3481 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     3541 tccattcgac caattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3601 ggccaatgcc ctggctcaca aataccactg agatccagac atgataagat acattgatga
     3661 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3721 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3781 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3841 cctctacaaa tgtggtatgg ctgattatga tccccaggaa gctcctctgt gtcctcataa
     3901 accctaacct cctctacttg agaggacatt ccaatcatag gctgcccatc caccctctgt
     3961 gtcctcctgt taattaggtc acttaacaaa aaggaaattg ggtaggggtt tttcacagac
     4021 cgctttctaa gggtaatttt aaaatatctg ggaagtccct tccactgctg tgttccagaa
     4081 gtgttggtaa acagcccaca aatgtcaaca gcagaaacat acaagctgtc agctttgcac
     4141 aagggcccaa caccctgctc atcaagaagc actgtggttg ctgtgttagt aatgtgcaaa
     4201 acaggaggca cattttcccc acctgtgtag gttccaaaat atctagtgtt ttcattttta
     4261 cttggatcag gaacccagca ctccactgga taagcattat ccttatccaa aacagccttg
     4321 tggtcagtgt tcatctgctg actgtcaact gtagcatttt ttggggttac agtttgagca
     4381 ggatatttgg tcctgtagtt tgctaacaca ccccaggtgg cacttttcgg ggaaatgtgc
     4441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5701 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6241 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6301 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6361 aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
