LOCUS       sequence-469796-        3039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses human alpha-synuclein with a 7-amino acid 
            insertion (MAAAEKT) in the pRK172 backbone for bacterial expression 
            in E. coli. The insertion corresponds to the JOS mutation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3039)
  AUTHORS   Yang Y, Garringer HJ, Shi Y, Lovestam S, Peak-Chew S, Zhang X, 
            Kotecha A, Bacioglu M, Koto A, Takao M, Spillantini MG, Ghetti B, 
            Vidal R, Murzin AG, Scheres SHW, Goedert M
  TITLE     New SNCA mutation and structures of alpha-synuclein filaments from 
            juvenile-onset synucleinopathy.
  JOURNAL   Acta Neuropathol. 2023 May;145(5):561-572. doi: 
            10.1007/s00401-023-02550-8. Epub 2023 Feb 27.
  PUBMED    36847833
REFERENCE   2  (bases 1 to 3039)
  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: 
            "10.1007/s00401-023-02550-8"; journalName: "Acta Neuropathol"; date:
            "2023-05"; volume: "145"; issue: "5"; pages: "561-572"
COMMENT     Sequence Label: 7AA-aSyn-pRK172
FEATURES             Location/Qualifiers
     source          1..3039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     197..216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(435..539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     607..625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(670..717)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     713..731
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1296..1318
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1349..1368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1350..1368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1421..1443)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1529..1668
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1584..1603
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1683..1700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1854..2442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1934..1953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2613..3039,1..365,366..434))
                     /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"
ORIGIN
        1 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
       61 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      181 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      541 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
      601 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattatccgg atatagttcc
      661 tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta
      721 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
      781 ggaattcaag cttggatgga acatctgtca gcagatctca agaaactggg agcaaagata
      841 tttcttaggc ttcaggttca tagtcttgat acccttcctc agaaggcatt tcataagcct
      901 cattgtcagg atccacaggc atatcttcca gaattccttc ctgtggggct ccttcttcat
      961 tcttgcccaa ctggtccttt ttgacaaagc cagtggctgc tgcaatgctc cctgctccct
     1021 ccactgtctt ctgggctact gctgtcacac ccgtcaccac tgctcctcca acatttgtca
     1081 cttgctcttt ggtcttctca gccactgttg ccacaccatg caccactccc tccttggttt
     1141 tggagcctac atagagaaca ccctcttttg tctttcctgc tgcttctgcc acaccctgtt
     1201 tagtcttttc tgctgctgcc atggttttct cagcagcagc cacaactccc tccttggcct
     1261 ttgaaagtcc tttcatgaat acatccatat gtatatctcc ttcttaaagt taaacaaaat
     1321 tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat
     1381 cgagatcctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1441 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1501 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     1561 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     1621 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     1681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2281 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     2941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagc
//
