LOCUS       sequence-469467-        4056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Tau-RING I18R-M72E mutant in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4056)
  AUTHORS   Miller LVC, Papa G, Vaysburd M, Cheng S, Sweeney PW, Smith A, Franco
            C, Katsinelos T, Huang M, Sanford SAI, Benn J, Farnsworth J, 
            Higginson K, Joyner H, McEwan WA, James LC
  TITLE     Co-opting templated aggregation to degrade pathogenic tau assemblies
            and improve motor function.
  JOURNAL   Cell. 2024 Oct 17;187(21):5967-5980.e17. doi: 
            10.1016/j.cell.2024.08.024. Epub 2024 Sep 13.
  PUBMED    39276772
REFERENCE   2  (bases 1 to 4056)
  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.cell.2024.08.024"; journalName: "Cell"; date: 
            "2024-10-17- 17"; volume: "187"; issue: "21"; pages: "5967-5980.e17"
COMMENT     Sequence Label: pOPTG-Tau-RING (M72E/I18R)
FEATURES             Location/Qualifiers
     source          1..4056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..54)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(208..796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(288..307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(967..1758,1759..1827))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1590..1609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1828..1932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2000..2018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2063..2110)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2106..2124
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3633..3650)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3664..3686
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3759..3778)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3760..3778)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3831..3853)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    join(3939..4056,1..22)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3994..4013
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
       61 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
      661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1261 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1561 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     1621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1981 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattatc
     2041 cggctatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     2101 gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt
     2161 tagcagccgg atcagcgcgc acgcgtggta ccttattcgc gtgcctcttg gctgatttct
     2221 ttcagattat taacctcatt ggccagctga cgattcggac gcagattttt cagcagaaaa
     2281 cgctgacgac aaaccggaca aacgctacca ccgcctttac caacctggct aatgcattct
     2341 tgacaaaagc tgtgaccaca ttcaatacta accggttcaa caaacgggtc cagacaacgc
     2401 ggacaggtaa cttcttccca catcatggtc agacgtgctg cgcttgccat acctgaaccg
     2461 ccaccgctac caaattccag accttgtttt gccaggcttg cgctaacttc atcggccagg
     2521 gttgccagct gcggactatc aaccatatca atgctaccgg tactgctaac attactcaga
     2581 tgacgcggac tggtatcacc gctaacaacc ggacttttat acacgatttc ggcaccatga
     2641 tcggttttgg cttttgcatt ttcgcggaag gtcagtttat gggtttcaat ctttttgtta
     2701 ccgccacctg gaacatgggt gatgttatcc agtgagccaa ttttgctctg gacgcgatct
     2761 ttgaaatcca gtttttcgct tttcacttca acctgaccac cacctggttt atgatgaata
     2821 ttacccagtg aaccgcattt gctggtaact ttgctcagat caaccggttt atacacaatc
     2881 tgaacgctgc caccaccgct cacatgttta atgttatctt tgctaccaca tttgctctgc
     2941 acattgctca ggtccagttt cttattaatg atctgaactt taccaccacc aggctgatgt
     3001 ttcaggttct cggtgctgcc gattttgctt ttaacatttt tcagatcagg catcggaacc
     3061 ggtgcggtct gcaggcgact ttttgcactg ctcgggcttt tcggaggggt acgaacaact
     3121 gcaacctttt tcggttcacg tgtaggcggt gtcggcaggc tcggtgtacg gctacggcta
     3181 cccggtgtac ccggagaacc cggactgcta taacctgaac gatcaccgct tttaggcggt
     3241 tcaccgctgc tcggaggtgt tttcggtgcc ggaggggttt ttgccggaat acgggttgca
     3301 tttgcctgac ctttctgacc aggcggtgct gcaccacgcg gtgttgcaat tttggttttg
     3361 ccatctgcac cttttgcctt tttatcatca ctaccggtgc catctttgct tttgctaacc
     3421 atacgagcct gggtaacatg acctgctgct tcatcttcca ggctcggggt atcgccaata
     3481 cctgcttctt cggctttcag acctgcatcg gtatcaccct cttgatcctg atgcatggta
     3541 taaccacctt gatctttacg atcacccaga ccataggtgc ctgcatgatc ttccataact
     3601 tcaaattctt gacgcggttc tgccatggtc atatggtgat ggtgatgatg agccatattt
     3661 atatctcctt cttaaagtta aacaaaatta tttctagagc tagcgaattc caattggagc
     3721 tctccggagc cggcactaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat
     3781 ttcgcgggat cgagatcctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3841 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3901 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3961 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     4021 tgcaccattg cggtgtgaaa taccgcacag atgcgt
//
