LOCUS       sequence-374444-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  K7A/D69A p62 double mutant (PB1AA). Internal reference: SMC532.
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 Double Mutation/Silent
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1327
                     /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(765..783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1033..1052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2808..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2845..2864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2899..2918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2936..3391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3073..3094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3285..3304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3418..3522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3524..3881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3524..3715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3547..3567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3732..3867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3916..4710
                     /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             3916..4710
                     /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(3970..3989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4580..4599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4898..4917)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4942..4989
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5318..5906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5807..5826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      661 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      781 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      841 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      961 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1021 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1081 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1141 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1201 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1261 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagctca aatggcgtcg ctcaccgtgg ctgcctacct
     1381 tctgggcaag gaggacgcgg cgcgcgagat tcgccgcttc agcttctgct gcagccccga
     1441 gcctgaggcg gaagccgagg ctgcggcggg tccgggaccc tgcgagcggc tgctgagccg
     1501 ggtggccgcc ctgttccccg cgctgcggcc tggcggcttc caggcgcact accgcgctga
     1561 ggacggggac ttggttgcct tttccagtga cgaggaattg acaatggcca tgtcctacgt
     1621 gaaggatgac atcttccgaa tctacattaa agagaaaaaa gagtgccggc gggaccaccg
     1681 cccaccgtgt gctcaggagg cgccccgcaa catggtgcac cccaatgtga tctgcgatgg
     1741 ctgcaatggg cctgtggtag gaacccgcta caagtgcagc gtctgcccag actacgactt
     1801 gtgtagcgtc tgcgagggaa agggcttgca ccgggggcac accaagctcg cattccccag
     1861 ccccttcggg cacctgtctg agggcttctc gcacagccgc tggctccgga aggtgaaaca
     1921 cggacacttc gggtggccag gatgggaaat gggtccacca ggaaactgga gcccacgtcc
     1981 tcctcgtgca ggggaggccc gccctggccc cacggcagaa tcagcttctg gtccatcgga
     2041 ggatccgagt gtgaatttcc tgaagaacgt tggggagagt gtggcagctg cccttagccc
     2101 tctgggcatt gaagttgata tcgatgtgga gcacggaggg aaaagaagcc gcctgacccc
     2161 cgtctctcca gagagttcca gcacagagga gaagagcagc tcacagccaa gcagctgctg
     2221 ctctgacccc agcaagccgg gtgggaatgt tgagggcgcc acgcagtctc tggcggagca
     2281 gatgaggaag atcgccttgg agtccgaggg gcgccctgag gagcaaatgg aatccgacaa
     2341 ctgttcagga ggagatgatg actggaccca tctgtcttca aaagaagtgg acccgtctac
     2401 aggtgaactc cagtccctac agatgccaga atccgaaggg ccaagctctc tggacccctc
     2461 ccaggaggga cccacagggc tgaaggaagc tgccttgtac ccacatctcc cgccagaggc
     2521 tgacccgcgg ctgattgagt ccctctccca gatgctgtcc atgggcttct ctgatgaagg
     2581 cggctggctc accaggctcc tgcagaccaa gaactatgac atcggagcgg ctctggacac
     2641 catccagtat tcaaagcatc ccccgccgtt gtgacccggg atccaccgga tctagataac
     2701 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2941 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3001 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3061 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3121 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3181 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3241 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3301 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3361 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3541 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3781 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3841 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3901 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3961 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4021 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4081 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4141 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4201 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4261 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4321 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4381 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4441 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4501 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4561 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4621 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4681 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4741 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4801 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4861 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4921 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4981 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5041 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5101 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5161 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5221 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5281 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5341 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5401 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5461 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5521 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5581 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5641 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5701 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5761 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5821 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5881 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5941 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6001 accgtattac cgc
//
