LOCUS       sequence-400879-        8861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Centromeric yeast expression vector for hDHDDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8861)
  AUTHORS   Galosi S, Edani BH, Martinelli S, Hansikova H, Eklund EA, Caputi C, 
            Masuelli L, Corsten-Janssen N, Srour M, Oegema R, Bosch DGM, Ellis 
            CA, Amlie-Wolf L, Accogli A, Atallah I, Averdunk L, Baranano KW, Bei
            R, Bagnasco I, Brusco A, Demarest S, Alaix AS, Di Bonaventura C, 
            Distelmaier F, Elmslie F, Gan-Or Z, Good JM, Gripp K, Kamsteeg EJ, 
            Macnamara E, Marcelis C, Mercier N, Peeden J, Pizzi S, Pannone L, 
            Shinawi M, Toro C, Verbeek NE, Venkateswaran S, Wheeler PG, 
            Zdrazilova L, Zhang R, Zorzi G, Guerrini R, Sessa WC, Lefeber DJ, 
            Tartaglia M, Hamdan FF, Grabinska KA, Leuzzi V
  TITLE     De novo DHDDS variants cause a neurodevelopmental and 
            neurodegenerative disorder with myoclonus.
  JOURNAL   Brain. 2022 Mar 29;145(1):208-223. doi: 10.1093/brain/awab299.
  PUBMED    34382076
REFERENCE   2  (bases 1 to 8861)
  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"; journalName: 
            "Brain"; date: "2022-03-29- 29"; volume: "145"; issue: "1"; pages: 
            "208-223"
COMMENT     Sequence Label: pTL-DHDDS
FEATURES             Location/Qualifiers
     source          1..8861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1650..1897
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1666..1684)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1920..1936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1920..1936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1933..1955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1944..1960
                     /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(1968..1974,1975..1992,1993..1998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2013..2034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2151..2168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2322..2910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2402..2421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3081..3872,3873..3941))
                     /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     3704..3723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3942..4046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4114..4132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             4695..5789
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYA
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVRHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     misc_feature    6584..7746
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      6584..7421
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     primer_bind     8488..8510
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8502..8519
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8503..8519
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttcgggggat ccgtttaaac tcctcaggaa catccgactt gtcaattttc aagtgatttc
       61 tttctgtcat gggaatttca gaaagtgctc ttgacaacaa caacactaac aagggacgta
      121 cttttttgcc ttctgtttca aaatagtaac tagtaacctt gtttagcacc ggatggccag
      181 acccgatgag ggcaactata tttttggcca atgtgttcat ctccttcgag actaatgata
      241 tgggattatt ccaaagaatc ttgggagtta ccaacttact ggcagcgttt aaagcaactg
      301 caaaggagga tttaaagcgg catcttcgag atgaaatcaa tttgatgtga tgagcagcgc
      361 cagacctttg aaacatcgtc ccactgtact caattcggac tgaatggcac acttcttgtg
      421 ttgtagaaaa tctctataga ttataatgtg tcttacttta atcttctaat cccctaaccc
      481 tgttaccctc gctcgggctt tttccgaaga tgccaacaaa atgcttagaa tgaaaaaaaa
      541 aaaaattttc gtaataatga caggtttcaa tagaacttaa gcagtaaaaa tagggtaaac
      601 acaggtaaaa gacggtatgt catggatcaa ggaaggagag ctgtcacttt gggagcggtt
      661 ctgtgccaac atcataaagg caggcccaat gccgaaacac attgcattca taatggacgg
      721 gaaccgtcgc tatgccaaga agtgccaggt ggagcggcag gaaggccact cacagggctt
      781 caacaagcta gctgagactc tgcggtggtg tttgaacctg ggcatcctag aggtgacagt
      841 ctacgcattc agcattgaga acttcaaacg ctccaagagt gaggtagacg ggcttatgga
      901 tctggcccgg cagaagttca gccgcttgat ggaagaaaag gagaaactgc agaagcatgg
      961 ggtgtgtatc cgggtcctgg gcgatctgca cttgttgccc ttggatctcc aggagctgat
     1021 tgcacaagct gtacaggcca cgaagaacta caacaagtgt ttcctgaatg tctgttttgc
     1081 atacacatcc cgtcatgaga tcagcaatgc tgtgagagag atggcctggg gggtggagca
     1141 aggcctgttg gatcccagtg atatctctga gtctctgctt gataagtgcc tctataccaa
     1201 ccgctctcct catcctgaca tcttgatacg gacttctgga gaagtgcggc tgagtgactt
     1261 cttgctatgg cagacctctc actcctgcct ggtgttccaa cccgttctgt ggccagagta
     1321 tacattttgg aacctcttcg aggccatcct gcagttccag atgaaccata gcatgcttca
     1381 gaaggcccga gacatgtatg cagaggagcg gaagaggcag cagctggaga gggaccaggc
     1441 tacagtgaca gagcagctgc tgcgagaggg gctccaagcc agtggggacg cccagctccg
     1501 aaggacacgc ttgcacaaac tctcggccag acgggaagag cgagtccaag gcttcctgca
     1561 ggccttggaa ctcaagcgag ctgactggct ggcccgtctg ggcactgcat cagcctgagc
     1621 ggccgctagg gccctgcagg agggccgcat catgtaatta gttatgtcac gcttacattc
     1681 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1741 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     1801 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     1861 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc caagcttggc gtaatcatgg
     1921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4081 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4141 cttcaagaat tagcttttca attcatcatt ttttttttat tctttttttt gatttcggtt
     4201 tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca
     4261 cagacttaga ttggtatata tacgcatatg gtgtgggaat actcaggtat cgtaagatgc
     4321 aagagttcga atctcttagc aaccattatt tttttcctca acataacgag aacacacagg
     4381 ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg
     4441 cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc
     4501 ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata
     4561 cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc
     4621 gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat atatttcaag
     4681 gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg
     4741 gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca
     4801 atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg
     4861 ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg
     4921 ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa
     4981 tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca tccgactctc
     5041 ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca
     5101 gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg
     5161 cttgggatag tgaacaatac gccgttccag aagtgcaaag aatcacaaga atggccgctt
     5221 tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatgttt
     5281 tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac gaattcccta
     5341 cattgaaggt tcgacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa
     5401 cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag
     5461 cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag
     5521 acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa
     5581 agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat
     5641 tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag
     5701 gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg
     5761 ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca
     5821 taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt
     5881 catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg
     5941 aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa
     6001 agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg
     6061 taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt tctctaaaaa
     6121 aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc
     6181 aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc
     6241 agaaacggcc ttaacgacgt accggatcat gcatgtatac taaactcaca aattagagct
     6301 tcaatttaat tatatcagtt attacccaat tctcatgttt gacagcttat catcgatcgt
     6361 ccaactgcat ggagatgagt cgtggcaaga ataccaagag ttcctcggtt tgccagttat
     6421 taaaagactc gtatttccaa aagactgcaa catactactc agtgcagctt cacagaaacc
     6481 tcattcgttt attcccttgt ttgattcaga agcaggtggg acaggtgaac ttttggattg
     6541 gaactcgatt tctgactggg ttggaaggca agagagcccc gagagcttac attttatgtt
     6601 agctggtgga ctgacgccag aaaatgttgg tgatgcgctt agattaaatg gcgttattgg
     6661 tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa gactctaaca aaatagcaaa
     6721 tttcgtcaaa aatgctaaga aataggttat tactgagtag tatttattta agtattgttt
     6781 gtgcacttgc ctgcaagcct tttgaaaagc aagcataaaa gatctaaaca taaaatctgt
     6841 aaaataacaa gatgtaaaga taatgctaaa tcatttggct ttttgattga ttgtacagga
     6901 aaatatacat cgcagggggt tgacttttac catttcaccg caatggaatc aaacttgttg
     6961 aagagaatgt tcacaggcgc atacgctaca atgacccgat tcttgctagc cttttctcgg
     7021 tcttgcaaac aaccgccggc agcttagtat ataaatacac atgtacatac ctctctccgt
     7081 atcctcgtaa tcattttctt gtatttatcg tcttttcgct gtaaaaactt tatcacactt
     7141 atctcaaata cacttattaa ccgcttttac tattatcttc tacgctgaca gtaatatcaa
     7201 acagtgacac atattaaaca cagtggtttc tttgcataaa caccatcagc ctcaagtcgt
     7261 caagtaaaga tttcgtgttc atgcagatag ataacaatct atatgttgat aattagcgtt
     7321 gcctcatcaa tgcgagatcc gtttaaccgg accctagtgc acttacccca cgttcggtcc
     7381 actgtgtgcc gaacatgctc cttcactatt ttaacatgtg gaattaattc tcatgtttga
     7441 cagcttatca tcgaactcta agaggtgata cttatttact gtaaaactgt gacgataaaa
     7501 ccggaaggaa gaataagaaa actcgaactg atctataatg cctattttct gtaaagagtt
     7561 taagctatga aagcctcggc attttggccg ctcctaggta gtgctttttt tccaaggaca
     7621 aaacagtttc tttttcttga gcaggtttta tgtttcggta atcataaaca ataaataaat
     7681 tatttcattt atgtttaaaa ataaaaaata aaaaagtatt ttaaattttt aaaaaagttg
     7741 attataagca tgtgaccttt tgcaagcaat taaattttgc aatttgtgat ttaggcaaaa
     7801 gttactattt ctggctcgtg taatatatgt atgctaatgt gaacttttac aaagtcgata
     7861 tggacttagt caaaagaaat tttcttaaaa atatatagca ctagccaatt tagcacttct
     7921 ttatgagata tattatagac tttattaagc cagatttgtg tattatatgt atttacccgg
     7981 cgaatcatgg acatacattc tgaaataggt aatattctct atggtgagac agcatagata
     8041 acctaggata caagttaaaa gctagtactg ttttgcagta atttttttct tttttataag
     8101 aatgttacca cctaaataag ttataaagtc aatagttaag tttgatattt gattgtaaaa
     8161 taccgtaata tatttgcatg atcaaaaggc tcaatgttga ctagccagca tgtcaaccac
     8221 tatattgatc accgatatta ggacttccac accaactagt aatatgacaa taaattcaag
     8281 atattcttca tgagaatggc ccagcgatta tgcggtgtga aataccgcac agatgcgtaa
     8341 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     8401 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     8461 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     8521 aattcttatt acgatcctcg cgccccctac ccaccgtact cgtcaattcc aagggcatcg
     8581 gtaaacatct gctcaaactc gaagtcggcc atatccagag cgccgtaggg ggcggagtcg
     8641 tggggggtaa atcccggacc cggggaatcc ccgtccccca acatgtccag atcgaaatcg
     8701 tctagcgcgt cggcatgcat gtgctctgta tgtatataaa actcttgttt tcttcttttc
     8761 tctaaatatt ctttccttat acattaggtc ctttgtagca taaattacta tacttctata
     8821 gacacgcaaa cacaaataca cacactaaat taccggatca a
//
