LOCUS       sequence-464210-        2958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of alpha synuclein A53T mutant, tagged with the
            final beta strand of mNeonGreen-3K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2958)
  AUTHORS   Sitron CS, Trinkaus VA, Galesic A, Garhammer M, Yuste-Checa P, 
            Dransfeld U, Feigenbutz D, Zhang J, Ivashko L, Dudanova I, Harper 
            JW, Hartl FU
  TITLE     alpha-Synuclein aggregates inhibit ESCRT-III through sequestration 
            and collateral degradation.
  JOURNAL   Mol Cell. 2025 Sep 18;85(18):3505-3523.e17. doi: 
            10.1016/j.molcel.2025.08.022. Epub 2025 Sep 10.
  PUBMED    40934925
REFERENCE   2  (bases 1 to 2958)
  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.molcel.2025.08.022"; journalName: "Mol Cell"; date: 
            "2025-09-18- 18"; volume: "85"; issue: "18"; pages: "3505-3523.e17"
COMMENT     Sequence Label: pT7-7 a-syn A53T D115A mNeonGreen-3K-B11
FEATURES             Location/Qualifiers
     source          1..2958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     757..774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1009..1031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1096..1115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1096..1114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1146..1168
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(join(1793..2584,2585..2653))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2416..2435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2654..2758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2826..2844
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      781 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      841 ggaagatctt ccggaagatc ttcctatggt gcactctcag tacaatctgc tctgatgcgc
      901 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgatgac gcgccatgac
      961 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1021 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcccagcgat tcgaacttct
     1081 gatagacttc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
     1141 ataattttgt ttaactttaa gaaggagata tacatatgga tgtattcatg aaaggacttt
     1201 caaaggccaa ggagggagtt gtggctgctg ctgagaaaac caaacagggt gtggcagaag
     1261 cagcaggaaa gacaaaagag ggtgttctct atgtaggctc caaaaccaag gagggagtgg
     1321 tgcatggtgt gacaacagtg gctgagaaga ccaaagagca agtgacaaat gttggaggag
     1381 cagtggtgac gggtgtgaca gcagtagccc agaagacagt ggagggagca gggagcattg
     1441 cagcagccac tggctttgtc aaaaaggacc agttgggcaa gaatgaagaa ggagccccac
     1501 aggaaggaat tctggaagcg atgcctgtgg atcctgacaa tgaggcttat gaaatgcctt
     1561 ctgaggaagg gtatcaagac tacgaacctg aagccggtag tggtggaagt ggtacggaat
     1621 tgaactttaa agaatggcaa aaagcgttta ccgacatgat gtaagaaata tctttgctcc
     1681 cagtttcttg agatctgctg acagatgttc catccaagct tatcgatgat aagctgtcaa
     1741 acatgagaat taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2401 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2761 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     2821 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga ataaaaggat ctaggtgaag
     2881 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2941 tcagaccccg tagaaaag
//
