LOCUS       sequence-171255-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human YOD1 C160S mutant with N-terminal GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5831)
  AUTHORS   Papadopoulos C, Kirchner P, Bug M, Grum D, Koerver L, Schulze N, 
            Poehler R, Dressler A, Fengler S, Arhzaouy K, Lux V, Ehrmann M, 
            Weihl CC, Meyer H
  TITLE     VCP/p97 cooperates with YOD1, UBXD1 and PLAA to drive clearance of 
            ruptured lysosomes by autophagy.
  JOURNAL   EMBO J. 2016 Oct 17. pii: e201695148.
  PUBMED    27753622
REFERENCE   2  (bases 1 to 5831)
  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: "EMBO J. 
            2016 Oct 17. pii: e201695148."
COMMENT     Sequence Label: pEGFP-YOD1-C160S
FEATURES             Location/Qualifiers
     source          1..5831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    745..769
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1814..1838)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2007..2128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2044..2063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2098..2117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2135..2590)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2272..2293)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2484..2503
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2617..2721
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2723..3080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2723..2914
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2746..2766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2931..3066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3115..3909
                     /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             3115..3909
                     /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(3169..3188)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3779..3798
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4097..4116)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4141..4188
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4517..5105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5006..5025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5280..5583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5584..5787
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5738..5758
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5826..5831,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctcgaggtta tcaaacaagt ttgtacaaaa aagttggcat gtttggcccc
      781 gctaaaggtc gccattttgg agtccacccg gcgcctggtt tccccggcgg cgtctcccaa
      841 caggctgccg ggaccaaagc tggccccgcg ggtgcctggc ctgtgggcag ccggaccgac
      901 acgatgtggc ggctccgctg caaggccaag gacggcaccc atgttttgca ggggctgtcc
      961 agccggaccc gggtgcggga actccagggc caaattgccg ccatcaccgg gatcgccccc
     1021 ggcggtcagc gaatcctcgt cggataccct cccgagtgcc tggatctcag caatggggat
     1081 accattctgg aagacttgcc catccaatct ggtgacatgc tgatcattga agaagaccaa
     1141 accaggccca gaagttcacc tgcatttact aaacgtggtg cttctagtta cgtcagggaa
     1201 actttgcctg tgcttaccag aaccgtggtc ccagcagaca actcttccct ctttactagt
     1261 gtgtactatg tcgtcgaagg aggagtcttg aatccagctt gtgcccctga gatgagacgc
     1321 ctcatagcac aaattgtagc aagcgatcca gacttctata gtgaggcaat actgggaaaa
     1381 acaaatcaag agtactgtga ctggatcaaa agggatgaca cttggggagg agcaatagag
     1441 atatcgattt tgtccaagtt ttaccaatgt gaaatatgtg tagtggatac acagacagta
     1501 agaattgatc gttttgggga agatgcagga tataccaaaa gggttctgct tatttatgat
     1561 ggcatccact atgatccact tcagcgtaac ttccctgatc cagatacacc tcctctgacc
     1621 attttctcct ctaatgatga tattgttctt gtacaagcac tggaattagc agatgaagct
     1681 agaagaagga gacagtttac tgatgtcaac cgcttcaccc tgagatgcat ggtatgtcag
     1741 aaaggattaa ctggacaagc agaagcaagg gaacatgcca aggagacagg ccataccaac
     1801 tttggagaag tgtgaccaac tttcttgtac aaagtggttc gataacgaat tctgcagtcg
     1861 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     1921 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1981 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2041 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2101 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     2161 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2221 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2281 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2341 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2401 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2461 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2521 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2581 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2641 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2701 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3001 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3061 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3121 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3181 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3241 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3301 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3361 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3421 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3481 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3541 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3601 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3661 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3721 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3781 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3841 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3901 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3961 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4021 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4081 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4141 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4201 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4261 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4321 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4381 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4441 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4501 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4561 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4621 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4681 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4741 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4801 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4861 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4921 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4981 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5041 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5101 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5161 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5221 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5281 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5341 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5401 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5461 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5521 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5581 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5641 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5701 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5761 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     5821 cggtcgccac c
//
