LOCUS       sequence-430535-        5464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the endolysosomal damage marker Galectin-3 with an 
            N-terminal mRuby3 tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5464)
  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 5464)
  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 mRuby3-Galectin-3
FEATURES             Location/Qualifiers
     source          1..5464
                     /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      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             667..1377
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     polyA_signal    2306..2427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2343..2362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2397..2416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2434..2889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2571..2592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2783..2802
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2916..3020
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3022..3379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3022..3213
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3045..3065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3230..3365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3414..4208
                     /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             3414..4208
                     /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(3468..3487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4078..4097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4396..4415)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4440..4487
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4816..5404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5305..5324
                     /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 gccaccatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
      721 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcagaccg
      781 tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
      841 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cctttatcaa gtacccggcc
      901 gacatccctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
      961 agatacgaag atggtggagt cgtcaccgtc acgcaggaca ccagccttga ggatggcgag
     1021 ctcgtctaca acgtcaaggt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     1081 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     1141 agaggataca ctgacatcgc actgaaagtt gatggtggtg gccatctgca ctgcaacttc
     1201 gtgacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tgtccatgcc
     1261 gttgatcacc gcctggaaag gatcgaggag agtgacaatg aaacctacgt agtgcaaaga
     1321 gaagtggcag ttgccaaata cagcaacctt ggtggtggca tggacgagct gtacaagtcc
     1381 ggactcagat ctatggcaga caatttttcg ctccatgatg cgttatctgg gtctggaaac
     1441 ccaaaccctc aaggatggcc tggcgcatgg gggaaccagc ctgctggggc agggggctac
     1501 ccaggggctt cctatcctgg ggcctacccc gggcaggcac ccccaggggc ttatcctgga
     1561 caggcacctc caggcgccta ccctggagca cctggagctt atcccggagc acctgcacct
     1621 ggagtctacc cagggccacc cagcggccct ggggcctacc catcttctgg acagccaagt
     1681 gcccccggag cctaccctgc cactggcccc tatggcgccc ctgctgggcc actgattgtg
     1741 ccttataacc tgcctttgcc tgggggagtg gtgcctcgca tgctgataac aattctgggc
     1801 acggtgaagc ccaatgcaaa cagaattgct ttagatttcc aaagagggaa tgatgttgcc
     1861 ttccacttta acccacgctt caatgagaac aacaggagag tcattgtttg caatacaaag
     1921 ctggataata actggggaag ggaagaaaga cagtcagttt tcccatttga aagtgggaaa
     1981 ccattcaaaa tacaagtact ggttgaacct gaccacttca aggttgcagt gaatgatgct
     2041 cacttgttgc agtacaatca tcgggttaaa aaactcaatg aaatcagcaa actgggaatt
     2101 tctggtgaca tagacctcac cagtgcttca tataccatga tataagaatt ctgcagtcga
     2161 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac
     2221 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2281 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2341 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2401 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     2461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2881 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2941 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3001 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3061 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3121 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3361 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     3961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4201 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4381 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4441 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4501 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4561 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4621 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4681 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4921 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5461 catg
//
