LOCUS       sequence-443191-        7589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged human NOVA2 (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  AUTHORS   Mattioli F, Hayot G, Drouot N, Isidor B, Courraud J, Hinckelmann MV,
            Mau-Them FT, Sellier C, Goldman A, Telegrafi A, Boughton A, Gamble 
            C, Moutton S, Quartier A, Jean N, Van Ness P, Grotto S, Nambot S, 
            Douglas G, Si YC, Chelly J, Shad Z, Kaplan E, Dineen R, Golzio C, 
            Charlet-Berguerand N, Mandel JL, Piton A
  TITLE     De Novo Frameshift Variants in the Neuronal Splicing Factor NOVA2 
            Result in a Common C-Terminal Extension and Cause a Severe Form of 
            Neurodevelopmental Disorder.
  JOURNAL   Am J Hum Genet. 2020 Apr 2;106(4):438-452. doi: 
            10.1016/j.ajhg.2020.02.013. Epub 2020 Mar 19.
  PUBMED    32197073
REFERENCE   2  (bases 1 to 7589)
  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"; doi: 
            "10.1016/j.ajhg.2020.02.013"; journalName: "Am J Hum Genet"; date: 
            "2020-04-2- 2"; volume: "106"; issue: "4"; pages: "438-452"
COMMENT     Sequence Label: peGFP NOVA2 op
FEATURES             Location/Qualifiers
     source          1..7589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      899..908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(905..907,908..910,911..1621)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(905..907,908..910,911..1621)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(950..971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1625..1634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3180..3197)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3186..3410
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3456..3884
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3753..3774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3893..3913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3898..4227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4078..4213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4140..4159
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4294..5088
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4348..4367)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4958..4977
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5262..5383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5299..5318)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5353..5372
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5432..5448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5432..5448)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5445..5467)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5456..5472
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5480..5486,5487..5504,5505..5510))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5525..5546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5663..5680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5834..6422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5914..5933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6593..7384,7385..7453))
                     /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     7216..7235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7454..7558)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 gcttgccacc atggagcccg aggcccctga ttcacggaaa cgcccactgg aaacaccccc
     1681 tgaggtcgtc tgcactaaaa ggtcaaatac tggagaagag ggcgaatact tcctgaaggt
     1741 gctgatccct tcatatgccg ctgggagcat cattggaaag ggcgggcaga ccatcgtgca
     1801 gctgcagaag gagacaggcg ccaccatcaa gctgagtaag tcaaaagact tctaccccgg
     1861 gaccacagag cgagtgtgcc tggtccaggg aaccgctgaa gcactgaacg ccgtgcacag
     1921 ctttatcgct gagaaggtca gagaaattcc ccaggccatg acaaaacctg aggtggtcaa
     1981 catcctgcag ccacagacta ccatgaatcc cgatcgggct aagcaggcaa aactgattgt
     2041 gcccaactcc accgccggcc tgatcattgg gaagggaggc gctacagtga aagcagtcat
     2101 ggaacagagt ggcgcctggg tgcagctgtc acagaagccc gaggggatca atctgcagga
     2161 acgagtggtc accgtgagcg gagagcctga acaggtgcat aaggccgtga gcgccatcgt
     2221 gcagaaagtc caggaggacc cacagagctc ctcttgtctg aacatttcct acgcaaatgt
     2281 ggccggaccc gtcgccaact ctaatcctac aggcagccca tatgcttccc cagcagatgt
     2341 gctgcctgca gcagctgcag catccgctgc agcagcttct ggactgctgg gaccagcagg
     2401 gctggctgga gtgggagcct tccctgcagc cctgccagct ttttctggaa cagacctgct
     2461 ggccatcagt actgctctga acaccctggc cagctacggc tataacacta attccctggg
     2521 actgggcctg aacagcgccg cagcaagtgg cgtgctggct gcagtcgcag ctggagcaaa
     2581 cccagcagca gctgcagccg ctaatctgct ggcatcctac gcaggagagg caggagctgg
     2641 acctgcaggg ggagcagccc cccctccacc ccctccaccc ggagcactgg gctctttcgc
     2701 tctggctgca gccgctaatg gctatctggg agcaggagca ggaggaggag caggaggggg
     2761 aggcgggcca ctggtggcag ccgctgcagc agctggagca gcaggaggct ttctgaccgc
     2821 agagaagctg gctgcagaaa gcgccaaaga gctggtggaa atcgccgtcc ccgagaacct
     2881 ggtgggagcc attctgggca aggggggaaa aacactggtc gagtaccagg aactgactgg
     2941 ggccaggatc cagattagta agaaagggga atttctgcct ggaactcgca atcggagagt
     3001 gacaatcact ggcagcccag cagctaccca ggcagcacag tatctgattt cacagagagt
     3061 cacatacgaa cagggcgtgc gggcaagcaa tccccagaag gtcggctgag aattctgcag
     3121 atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     3181 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3241 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3301 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3361 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3421 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3601 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3661 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3781 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3841 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4141 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4201 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4261 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4321 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4381 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4441 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4501 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4561 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4621 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4681 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4741 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4801 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4861 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4921 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4981 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5041 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5101 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5161 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5221 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5401 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5461 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5521 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5581 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5641 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6241 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7561 catttccccg aaaagtgcca cctgacgtc
//
