LOCUS       sequence-171258-        7110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human p97 E578Q mutant with N-terminal GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7110)
  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 7110)
  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-p97-E578Q
FEATURES             Location/Qualifiers
     source          1..7110
                     /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"
     polyA_signal    3286..3407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3323..3342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3377..3396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3414..3869)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3551..3572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3763..3782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3896..4000
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4002..4359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4002..4193
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4025..4045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4210..4345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4394..5188
                     /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             4394..5188
                     /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(4448..4467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5058..5077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5376..5395)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5420..5467
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5796..6384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6285..6304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6559..6862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6863..7066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7017..7037
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7105..7110,1..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 ctcgagccat ggcctctgga gccgattcaa aaggtgatga tttatcaaca
      781 gccattctca aacagaagaa ccgtcccaat cggttaattg ttgatgaagc catcaatgaa
      841 gataacagtg tggtgtcctt gtcccagccc aagatggatg aactacagtt gttcagaggt
      901 gacacggtgt tgctaaaagg aaagaagaga agggaagctg tatgcattgt tctttctgat
      961 gacacgtgtt ctgatgagaa gattcgaatg aatagagttg ttcggaataa cctccgagtt
     1021 cgcctaggag atgtcatcag catccagcca tgccctgatg taaagtatgg caaacgtatc
     1081 catgtgctac ccattgatga cacagtggaa ggcatcactg gcaatctttt tgaggtatac
     1141 cttaagccgt acttcctgga agcatatcgg cccatccgta aaggagatat tttccttgtc
     1201 cggggtggga tgcgtgctgt ggagttcaaa gtagtagaga cagatcccag cccttactgt
     1261 attgttgctc cagacacagt gatccactgt gagggggagc caatcaagcg agaggatgag
     1321 gaggagtcct taaatgaagt aggctatgat gacatcggtg gttgcaggaa gcagctagct
     1381 cagataaagg agatggtgga gctgccactg agacatcctg cactctttaa ggcaattggt
     1441 gtgaagcctc ctcggggaat cttgctatat ggacctcctg ggacagggaa aaccttgatt
     1501 gcccgagctg tggcaaatga aactggagcc ttcttctttc tgatcaatgg tcctgaaatc
     1561 atgagcaaat tggctggtga gtctgagagc aaccttcgta aagcctttga ggaagctgaa
     1621 aagaatgctc ctgccatcat cttcatcgac gagcttgatg ccattgcacc caaaagagag
     1681 aaaactcacg gggaagtgga gcgtcgcatc gtgtctcagt tgttgaccct tatggatggc
     1741 ctaaagcaga gagcacatgt gatagttatg gcagcaacca atagacccaa cagcattgac
     1801 ccagccctac ggcgatttgg tcgctttgac agagaggtag atattggaat ccctgatgct
     1861 acaggacgtt tggaaattct tcagatccat accaagaaca tgaaactggc agatgatgtg
     1921 gacttggaac aggtagccaa tgagactcat ggtcatgtgg gtgctgactt ggcagccctg
     1981 tgttcagagg ctgctctaca ggccatccgg aaaaaaatgg acctcattga cctagaagat
     2041 gagaccattg acgctgaggt catgaattcc ctggcagtta ctatggatga cttccggtgg
     2101 gccttaagtc aaagcaaccc atcagcactt cgggaaactg tggtagaagt gccacaagta
     2161 acctgggaag acattggagg cctggaggat gtcaaacggg agcttcagga gttggttcag
     2221 tatcctgtgg agcatccaga caaattcctc aaatttggca tgactccttc caaaggtgtt
     2281 cttttctatg gaccgcctgg ctgtgggaaa accttactgg ccaaagccat tgctaatgaa
     2341 tgccaggcta acttcatctc catcaagggt cctgagctgc ttaccatgtg gtttggggaa
     2401 tctgaggcca atgtcaggga aatatttgac aaggcacgac aagctgcccc ctgtgtactc
     2461 ttctttgatc agttagattc aattgccaag gctcgtggtg gtaatattgg agatggtggt
     2521 ggagctgcag accgagtcat caatcagatc ctgacagaaa tggatggcat gtccacaaaa
     2581 aagaatgtgt ttatcattgg agctaccaac cggcctgaca tcattgatcc tgctatccta
     2641 agacctggcc gtcttgatca gctcatttat atcccacttc ctgatgagaa gtcccgtgtt
     2701 gccatcctaa aagccaatct gcgaaaatcc ccagttgcca aggatgtgga tttggagttc
     2761 ttggctaaga tgactaatgg cttttctgga gctgacctga cagaaatttg ccaacgtgct
     2821 tgtaaactag ccattcgtga atctatcgag agtgagatta ggcgagaacg agagaggcag
     2881 acaaatccat cagctatgga agtagaagag gatgatccag tgcctgagat ccgcagagat
     2941 cactttgagg aagccatgcg ttttgcccga cgttctgtca gtgataatga cattcggaag
     3001 tatgaaatgt ttgcccagac actgcagcaa agtcgaggtt ttggcagctt cagattccct
     3061 tcagggaacc agggtggagc tggcccaagt cagggcagtg gaggtggcac aggtggcaat
     3121 gtgtacacag aagacaatga cgatgacctc tatggcggat ccaccggatc tagataactg
     3181 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3241 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3301 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3361 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3421 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3481 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3541 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3601 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3661 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3721 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3781 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3841 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3901 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3961 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4381 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4441 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4501 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4561 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4621 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4681 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4741 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4801 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4861 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4921 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4981 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5041 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5101 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5161 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5221 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5281 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5341 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5401 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5461 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5521 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5581 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5641 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5701 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5941 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6481 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     6541 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     6601 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     6661 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     6721 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     6781 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     6841 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     6901 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     6961 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     7021 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     7081 tcagatccgc tagcgctacc ggtcgccacc
//
