LOCUS       sequence-374445-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the expression of a p62 mutant (LIR mutant: p62 3xDW + 
            silent mutation). Internal reference: SMC528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Wurzer B, Zaffagnini G, Fracchiolla D, Turco E, Abert C, Romanov J, 
            Martens S
  TITLE     Oligomerization of p62 allows for selection of ubiquitinated cargo 
            and isolation membrane during selective autophagy.
  JOURNAL   Elife. 2015 Sep 28;4:e08941. doi: 10.7554/eLife.08941.
  PUBMED    26413874
REFERENCE   2  (bases 1 to 6013)
  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"; journalName: "Elife."; 
            date: "2015-09-28"; volume: "4:e08941. doi"; pages: " 
            10.7554/eLife.08941"
COMMENT     Sequence Label: mCherry-p62-LIR-mutant (LIRAAAA)
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             625..1332
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(770..788)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1038..1057
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2813..2934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2850..2869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2904..2923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2941..3396)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3078..3099)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3290..3309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3423..3527
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3529..3886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3529..3720
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3552..3572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3737..3872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3921..4715
                     /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             3921..4715
                     /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(3975..3994)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4585..4604
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4903..4922)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4947..4994
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5323..5911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5812..5831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc
      661 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      721 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
      781 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
      841 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
      901 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      961 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1021 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1081 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1141 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1201 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1261 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1321 gagctgtaca agtccggact cagatctcga gctcaaatgg cgtcgctcac cgtgaaggcc
     1381 taccttctgg gcaaggagga cgcggcgcgc gagattcgcc gcttcagctt ctgctgcagc
     1441 cccgagcctg aggcggaagc cgaggctgcg gcgggtccgg gaccctgcga gcggctgctg
     1501 agccgggtgg ccgccctgtt ccccgcgctg cggcctggcg gcttccaggc gcactaccgc
     1561 gatgaggacg gggacttggt tgccttttcc agtgacgagg aattgacaat ggccatgtcc
     1621 tacgtgaagg atgacatctt ccgaatctac attaaagaga aaaaagagtg ccggcgggac
     1681 caccgcccac cgtgtgctca ggaggcgccc cgcaacatgg tgcaccccaa tgtgatctgc
     1741 gatggctgca atgggcctgt ggtaggaacc cgctacaagt gcagcgtctg cccagactac
     1801 gacttgtgta gcgtctgcga gggaaagggc ttgcaccggg ggcacaccaa gctcgcattc
     1861 cccagcccct tcgggcacct gtctgagggc ttctcgcaca gccgctggct ccggaaggtg
     1921 aaacacggac acttcgggtg gccaggatgg gaaatgggtc caccaggaaa ctggagccca
     1981 cgtcctcctc gtgcagggga ggcccgccct ggccccacgg cagaatcagc ttctggtcca
     2041 tcggaggatc cgagtgtgaa tttcctgaag aacgttgggg agagtgtggc agctgccctt
     2101 agccctctgg gcattgaagt tgatatcgat gtggagcacg gagggaaaag aagccgcctg
     2161 acccccgtct ctccagagag ttccagcaca gaggagaaga gcagctcaca gccaagcagc
     2221 tgctgctctg accccagcaa gccgggtggg aatgttgagg gcgccacgca gtctctggcg
     2281 gagcagatga ggaagatcgc cttggagtcc gaggggcgcc ctgaggagca aatggaatcc
     2341 gacaactgtt caggaggagc tgctgccgcg acccatctgt cttcaaaaga agtggacccg
     2401 tctacaggtg aactccagtc cctacagatg ccagaatccg aagggccaag ctctctggac
     2461 ccctcccagg agggacccac agggctgaag gaagctgcct tgtacccaca tctcccgcca
     2521 gaggctgacc cgcggctgat tgagtccctc tcccagatgc tgtccatggg cttctctgat
     2581 gaaggcggct ggctcaccag gctcctgcag accaagaact atgacatcgg agcggctctg
     2641 gacaccatcc agtattcaaa gcatcccccg ccgttgtgac ccgggatcca ccggatctag
     2701 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2761 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2941 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3001 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3061 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3121 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3181 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3241 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3301 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3361 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3541 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3721 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3781 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3841 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3901 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3961 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4021 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4081 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4141 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4201 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4261 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4321 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4381 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4441 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4501 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4561 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4621 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4681 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4741 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4801 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4861 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4921 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4981 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5041 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5101 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5161 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5221 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5461 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5581 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5641 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5701 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5761 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5821 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5881 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5941 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6001 ggataaccgt att
//
