LOCUS       sequence-177181-        5831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human YOD1 I292Q V295Q 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-MutS2
FEATURES             Location/Qualifiers
     source          1..5831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1297..1321
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2366..2390)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2559..2680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2596..2615)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2650..2669
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2687..3142)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2824..2845)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3036..3055
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3169..3273
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3275..3632
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3275..3466
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3298..3318)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3483..3618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3667..4461
                     /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             3667..4461
                     /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(3721..3740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4331..4350
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4649..4668)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4693..4740
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5069..5657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5558..5577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgaggt tatcaaacaa gtttgtacaa aaaagttggc
     1321 atgtttggcc ccgctaaagg tcgccatttt ggagtccacc cggcgcctgg tttccccggc
     1381 ggcgtctccc aacaggctgc cgggaccaaa gctggccccg cgggtgcctg gcctgtgggc
     1441 agccggaccg acacgatgtg gcggctccgc tgcaaggcca aggacggcac ccatgttttg
     1501 caggggctgt ccagccggac ccgggtgcgg gaactccagg gccaaattgc cgccatcacc
     1561 gggatcgccc ccggcggtca gcgaatcctc gtcggatacc ctcccgagtg cctggatctc
     1621 agcaatgggg ataccattct ggaagacttg cccatccaat ctggtgacat gctgatcatt
     1681 gaagaagacc aaaccaggcc cagaagttca cctgcattta ctaaacgtgg tgcttctagt
     1741 tacgtcaggg aaactttgcc tgtgcttacc agaaccgtgg tcccagcaga caactcttgc
     1801 ctctttacta gtgtgtacta tgtcgtcgaa ggaggagtct tgaatccagc ttgtgcccct
     1861 gagatgagac gcctcatagc acaaattgta gcaagcgatc cagacttcta tagtgaggca
     1921 atactgggaa aaacaaatca agagtactgt gactggatca aaagggatga cacttgggga
     1981 ggagcaatag agatatcgat tttgtccaag ttttaccaat gtgaaatatg tgtagtggat
     2041 acacagacag taagaattga tcgttttggg gaagatgcag gatataccaa aagggttctg
     2101 cttatttatg atggcatcca ctatgatcca cttcagcgta acttccctga tccagataca
     2161 cctcctctga ccattttctc ctctaatgat gatcaggttc ttcagcaagc actggaatta
     2221 gcagatgaag ctagaagaag gagacagttt actgatgtca accgcttcac cctgagatgc
     2281 atggtatgtc agaaaggatt aactggacaa gcagaagcaa gggaacatgc caaggagaca
     2341 ggccatacca actttggaga agtgtgacca actttcttgt acaaagtggt tcgataacga
     2401 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     2461 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2521 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2641 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2701 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2761 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2821 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2881 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2941 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3001 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3061 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3121 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3181 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3241 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3301 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3361 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3421 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3541 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3601 gaggcttwtt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3661 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3721 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3781 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3841 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3901 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3961 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4021 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4081 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4141 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4201 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4261 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4321 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4381 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4441 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4501 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4561 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4621 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4681 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4741 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4801 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4861 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4921 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4981 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5041 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5101 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5161 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5221 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5281 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5341 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5401 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5461 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5521 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5581 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5641 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5701 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5761 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     5821 atggagttcc g
//
