LOCUS       sequence-171124-        8456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of VCPIP1 with N-terminal GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8456)
  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 8456)
  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-VCPIP1
FEATURES             Location/Qualifiers
     source          1..8456
                     /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(4439..4463)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4632..4753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4669..4688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4723..4742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4760..5215)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4897..4918)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5109..5128
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5242..5346
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5348..5705
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5348..5539
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5371..5391)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5556..5691
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5740..6534
                     /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             5740..6534
                     /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(5794..5813)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6404..6423
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6722..6741)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6766..6813
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7142..7730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7631..7650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7905..8208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8209..8412
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8363..8383
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8451..8456,1..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 gtctcagccg
      781 ccgccgccgc cgcctccgtt gccgccgcca cctcctcccc ctgaggctcc acagactccg
      841 tcgtccttgg cgtcggcggc tgcttcgggg gggcttttga agcggagaga ccggagaatc
      901 ctttccggga gctgcccgga tccgaagtgt caggcgcgtc tatttttccc ggcctccggt
      961 tctgtcagca tcgagtgtac cgagtgcggc cagcggcacg agcagcaaca gctgctgggg
     1021 gttgaggagg tgaccgaccc ggacgtagtg ctacacaacc tgctgcggaa cgcgctgctc
     1081 ggggttacgg gggcacccaa gaagaacacg gaactggtaa aggtgatggg cctttccaac
     1141 tatcactgca aattgttgtc gcccatatta gctcgctatg gaatggacaa acagacaggc
     1201 cgggccaagc ttctccggga catgaaccag ggcgaactgt tcgattgcgc cttactgggt
     1261 gaccgcgcct tcctcataga accagagcat gttaacactg tgggctatgg caaggaccgc
     1321 tccggaagcc tcctgtattt gcatgacact ctggaggaca ttaagcgggc caataaaagc
     1381 caggaatgtc tcattccagt gcatgtggac ggggatggac actgcttggt gcatgctgtg
     1441 tctcgggctc tagtaggccg agagctcttc tggcatgcct taagagagaa tcttaaacag
     1501 cactttcagc agcacctggc ccgatatcaa gctctgttcc atgacttcat tgatgctgct
     1561 gagtgggagg acattatcaa tgagtgtgac cctctgtttg taccacctga gggtgttccc
     1621 ttgggcctga ggaatatcca catatttggt cttgccaatg tgctacatcg tcctattatt
     1681 ctgttagatt ccctcagtgg catgagaagc tctggtgatt attcagccac ctttctacct
     1741 gggctcatcc ctgcagagaa gtgcactggg aaagatggtc atttgaacaa accaatctgt
     1801 attgcatgga gcagctccgg tagaaaccat tatatcccct tggtaggcat aaaaggggct
     1861 gctttgccca aactgcctat gaatttgctt cctaaagcat ggggtgtgcc tcaggacctt
     1921 attaaaaagt acataaaact tgaagaggat ggtggttgtg ttattggagg tgacagaagt
     1981 ttgcaagata aatacttact taggcttgtt gctgctatgg aagaagtctt tatggacaaa
     2041 catggtatcc atcctagttt ggttgctgat gtccatcagt atttctacag aaggactgga
     2101 gtgataggag ttcagcctga ggaagtcaca gcagctgcta aaaaagcagt aatggataat
     2161 cgccttcaca aatgtttgct ctgtggtgcc ctttctgaac ttcatgttcc tccagagtgg
     2221 ttggctcctg gagggaaatt gtataacctg gcaaaaagta ctcatggaca gctgaggact
     2281 gacaaaaatt acagctttcc cttgaacaat ttggtttgct catatgattc agtgaaagat
     2341 gttctggtac cagactatgg aatgagtaac ctaacagctt gtaattggtg ccatggcaca
     2401 tctgtgcgaa aggtcagagg agatgggtct attgtgtatt tggatggaga cagaactaat
     2461 tctaggtcca ctggtggcaa atgtggttgt ggattcaaac acttttggga tggtaaggag
     2521 tatgacaatc taccagaagc tttccctatt actttagaat ggggtggaag agtggtcaga
     2581 gaaacagtat attggttcca gtatgaaagt gattcatctt tgaatagtaa tgtttacgat
     2641 gttgcaatga aacttgttac caagcacttt ccaggtgaat ttgggagtga aatcctagtt
     2701 cagaaagttg tccacactat attgcatcag actgccaaaa agaatcccga tgattatact
     2761 cctgtaaata tagatggtgc tcacgcccaa agagttggag atgttcaagg acaagaatca
     2821 gagtctcagc tcccaactaa aattattctt actggacaga aaacaaaaac tttgcacaag
     2881 gaggagttaa acatgagtaa aactgaaaga actattcaac agaatattac ggaacaggct
     2941 tctgtaatgc agaaacggaa aacagagaag ttaaaacaag aacaaaaagg gcaacccagg
     3001 actgtttctc ccagtaccat tcgtgatggt ccatcctctg cacctgctac acctaccaag
     3061 gctccctatt caccgacaac ttctaaggag aagaagatcc gaatcacaac taatgatgga
     3121 cgacagtcca tggttaccct taagtcttca acaacctttt ttgaacttca ggaaagtata
     3181 gccagagaat tcaacattcc tccatattta cagtgtattc gatacgggtt tcctcctaaa
     3241 gagttaatgc caccacaggc aggaatggaa aaggaaccag ttcctttaca gcatggcgac
     3301 agaattacaa tagaaattct aaaaagtaaa gctgaaggtg gtcagtctgc tgcagcacac
     3361 tcagcccaca ctgtgaaaca agaagatatt gctgttactg gtaaactgtc atctaaggaa
     3421 cttcaggagc aagctgaaaa agaaatgtac tccttgtgtc ttttagcaac attaatggga
     3481 gaagatgtat ggtcttatgc aaagggactt cctcacatgt ttcagcaggg tggtgtattc
     3541 tacagtatta tgaagaaaac catgggtatg gctgatggca agcattgtac ttttccacat
     3601 ctgcctggca aaacctttgt ctataatgct tctgaagata gactggaatt gtgtgtggat
     3661 gctgcaggac atttccccat tggtcctgat gttgaagatt tagttaaaga ggctgtaagt
     3721 caggttcgag cagaggctac tacaagaagt agggaatcaa gtccctcaca tgggctatta
     3781 aaactaggta gtggtggagt agtgaaaaag aaatctgagc aacttcataa cgtaactgcc
     3841 tttcagggaa aagggcattc tttaggaact gcatctggta acccacacct tgatccaaga
     3901 gctagggaaa cttcagttgt aagaaagcat aatacaggga cagactttag taatagttcc
     3961 actaaaacag agccttctgt attcacagct tcttctagta atagtgagct tattcgaata
     4021 gctcctggag tagtaacaat gagagacggc aggcagcttg atcctgattt ggttgaggcc
     4081 cagcgaaaaa aattgcagga aatggtttct tctattcagg cttcaatgga caggcacctt
     4141 cgggatcaaa gtacagagca gtcaccatct gatcttcctc aaaggaaaac agaagttgtg
     4201 agttcttctg caaagtctgg gagtcttcag actggtttgc ctgaatcttt tcctttaact
     4261 ggtggtactg aaaatttgaa tacagaaaca actgatggct gtgtagcaga tgcactggga
     4321 gcagcctttg ccacaaggtc aaaagcacaa aggggaaatt ccgtggagga gcttgaagag
     4381 atggatagtc aagatgctga gatgactaac acaactgagc caatggatca ctcttgatac
     4441 ccaactttct tgtacaaagt ggttcgataa cgaattctgc agtcgacggt accgcgggcc
     4501 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     4561 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4621 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4741 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4801 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4861 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4921 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4981 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5041 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5101 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5161 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5221 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5281 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5341 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5521 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5581 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5641 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5701 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5761 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5821 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5881 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5941 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6001 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6061 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6121 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6181 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6241 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6301 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6361 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6421 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6481 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6541 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6601 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6661 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     6721 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     6781 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6841 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6901 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6961 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7021 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7141 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7201 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7261 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7321 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7381 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7441 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7501 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7561 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7621 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7681 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7741 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7801 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     7861 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7921 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7981 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     8041 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     8101 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     8161 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     8221 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     8281 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     8341 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     8401 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccacc
//
