LOCUS       sequence-440345-        5149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the A53T mutant of alpha-synuclein with a C-terminal 
            mRuby3 tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5149)
  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 5149)
  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: pCMV-SNCA-A53T-mRuby3
FEATURES             Location/Qualifiers
     source          1..5149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1120..1827
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     polyA_signal    1991..2112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2028..2047)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2082..2101
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2119..2574)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2256..2277)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2468..2487
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2601..2705
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2707..3064
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2707..2898
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2730..2750)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2915..3050
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3099..3893
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3099..3893
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3153..3172)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3763..3782
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4081..4100)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4125..4172
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4501..5089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4990..5009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      661 gccaccgcca ccatggatgt attcatgaaa ggactttcaa aggccaagga gggagttgtg
      721 gctgctgctg agaaaaccaa acagggtgtg gcagaagcag caggaaagac aaaagagggt
      781 gttctctatg taggctccaa aaccaaggag ggagtggtgc atggtgtgac aacagtggct
      841 gagaagacca aagagcaagt gacaaatgtt ggaggagcag tggtgacggg tgtgacagca
      901 gtagcccaga agacagtgga gggagcaggg agcattgcag cagccactgg ctttgtcaaa
      961 aaggaccagt tgggcaagaa tgaagaagga gccccacagg aaggaattct ggaagatatg
     1021 cctgtggatc ctgacaatga ggcttatgaa atgccttctg aggaagggta tcaagactac
     1081 gaacctgaag ccggaggttc aggttcaggt ggaggatccg tgtctaaggg cgaagagctg
     1141 atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc
     1201 aaatgcacag gtgaaggaga aggcagaccg tacgagggag tgcaaaccat gaggatcaaa
     1261 gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat
     1321 ggcagccgta cctttatcaa gtacccggcc gacatccctg atttctttaa acagtccttt
     1381 cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc
     1441 acgcaggaca ccagccttga ggatggcgag ctcgtctaca acgtcaaggt cagaggggta
     1501 aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca
     1561 gagatgatgt atccagcaga tggtggtctg agaggataca ctgacatcgc actgaaagtt
     1621 gatggtggtg gccatctgca ctgcaacttc gtgacaactt acaggtcaaa aaagaccgtc
     1681 gggaacatca agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatcgaggag
     1741 agtgacaatg aaacctacgt agtgcaaaga gaagtggcag ttgccaaata cagcaacctt
     1801 ggtggtggca tggacgagct gtacaagtaa gaattctgca gtcgacggta ccgcgggccc
     1861 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     1921 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1981 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2041 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2101 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2161 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2221 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2281 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2341 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2401 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2461 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2521 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2581 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2641 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2701 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2761 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2821 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2881 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2941 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3001 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3061 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3121 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3181 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3241 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3301 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3361 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3421 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3481 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3541 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3601 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3661 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3721 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3781 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3841 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3901 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3961 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4021 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4081 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4141 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4201 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4261 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4321 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatg
//
