LOCUS       sequence-259425-        7186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7186)
  AUTHORS   Tresse E, Salomons FA, Vesa J, Bott LC, Kimonis V, Yao TP, Dantuma 
            NP, Taylor JP
  TITLE     VCP/p97 is essential for maturation of ubiquitin-containing 
            autophagosomes and this function is impaired by mutations that cause
            IBMPFD.
  JOURNAL   Autophagy. 2010 Feb 22. 6(2):.
  PUBMED    20104022
REFERENCE   2  (bases 1 to 7186)
  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: "Autophagy. 
            2010 Feb 22. 6(2):."
COMMENT     Sequence Label: VCP(A232E)-EGFP
FEATURES             Location/Qualifiers
     source          1..7186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2706..2715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2712..2714,2715..2717,2718..3425)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(2712..2714,2715..2717,2718..3425)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2757..2778)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3018..3037)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3365..3386
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3435..3452
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3456..3482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3610..3731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3647..3666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3701..3720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3738..4193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3875..3896)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4087..4106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4220..4324
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4326..4683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4326..4517
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4349..4369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4534..4669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4718..5512
                     /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             4718..5512
                     /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(4772..4791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5382..5401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5700..5719)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5744..5791
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6120..6708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6609..6628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6883..7186
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttgcgcgcg ccatggcttc tggagccgat tcaaaaggtg
      301 atgacctatc aacagccatt ctcaaacaga agaaccgtcc caatcggtta attgttgatg
      361 aagccatcaa tgaggacaac agtgtggtgt ccttgtccca gcccaagatg gatgaattgc
      421 agttgttccg aggtgacaca gtgttgctga aaggaaagaa gagacgagaa gctgtttgca
      481 tcgtcctttc tgatgatact tgttctgatg agaagattcg gatgaataga gttgttcgga
      541 ataaccttcg tgtacgccta ggggatgtca tcagcatcca gccatgccct gatgtgaagt
      601 acggcaaacg tatccatgtg ctgcccattg atgacacagt ggaaggcatt actggtaatc
      661 tcttcgaggt ataccttaag ccgtacttcc tggaagcgta tcgacccatc cggaaaggag
      721 acatttttct tgtccgtggt gggatgcgtg ctgtggagtt caaagtggtg gaaacagatc
      781 ctagccctta ttgcattgtt gctccagaca cagtgatcca ctgcgaaggg gagcctatca
      841 aacgagagga tgaggaagag tccttgaatg aagtagggta tgatgacatt ggtggctgca
      901 ggaagcagct agctcagata aaggagatgg tggaactgcc cctgagacat cctgccctct
      961 ttaaggaaat tggtgtgaag cctcctagag gaatcctgct ttacggacct cctggaacag
     1021 gaaagaccct gattgctcga gctgtagcaa atgagactgg agccttcttc ttcttgatca
     1081 atggtcctga gatcatgagc aaattggctg gtgagtctga gagcaacctt cgtaaagcct
     1141 ttgaggaggc tgagaagaat gctcctgcca tcatcttcat tgatgagcta gatgccatcg
     1201 ctcccaaaag agagaaaact catggcgagg tggagcggcg cattgtatca cagttgttga
     1261 ccctcatgga tggcctaaag cagagggcac atgtgattgt tatggcagca accaacagac
     1321 ccaacagcat tgacccagct ctacggcgat ttggtcgctt tgacagggag gtagatattg
     1381 gaattcctga tgctacagga cgcttagaga ttcttcagat ccataccaag aacatgaagc
     1441 tggcagatga tgtggacctg gaacaggtag ccaatgagac tcacgggcat gtgggtgctg
     1501 acttagcagc cctgtgctca gaggctgctc tgcaagccat ccgcaagaag atggatctca
     1561 ttgacctaga ggatgagacc attgatgccg aggtcatgaa ctctctagca gttactatgg
     1621 atgacttccg gtgggccttg agccagagta acccatcagc actgcgggaa accgtggtag
     1681 aggtgccaca ggtaacctgg gaagacatcg ggggcctaga ggatgtcaaa cgtgagctac
     1741 aggagctggt ccagtatcct gtggagcacc cagacaaatt cctgaagttt ggcatgacac
     1801 cttccaaggg agttctgttc tatggacctc ctggctgtgg gaaaactttg ttggccaaag
     1861 ccattgctaa tgaatgccag gccaacttca tctccatcaa gggtcctgag ctgctcacca
     1921 tgtggtttgg ggagtctgag gccaatgtca gagaaatctt tgacaaggcc cgccaagctg
     1981 ccccctgtgt gctattcttt gatgagctgg attcgattgc caaggctcgt ggaggtaaca
     2041 ttggagatgg tggtggggct gctgaccgag tcatcaacca gatcctgaca gaaatggatg
     2101 gcatgtccac taaaaaaaat gtgttcatca ttggcgctac caaccggcct gacatcattg
     2161 atcctgccat cctcagacct ggccgtcttg atcagctcat ctacatccca cttcctgatg
     2221 agaagtcccg tgttgccatc ctcaaggcta acctgcgcaa gtccccagtt gccaaggatg
     2281 tggacttgga gttcctggct aaaatgacta atggcttctc tggagctgac ctgacagaga
     2341 tttgccagcg tgcttgcaag ctggccatcc gtgaatccat cgagagtgag attaggcgag
     2401 aacgagagag gcagacaaac ccatcagcca tggaggtaga agaggatgat ccagtgcctg
     2461 agatccgtcg agatcacttt gaagaagcca tgcgctttgc gcgccgttct gtcagtgaca
     2521 atgacattcg gaagtatgag atgtttgccc agacccttca gcagagtcgg ggctttggca
     2581 gcttcagatt cccttcaggg aaccagggtg gagctggccc cagtcagggc agtggaggcg
     2641 gcacaggtgg cagtgtatac acagaagaca atgatgatga cctgtatggc tcggatccac
     2701 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2761 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2821 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2881 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2941 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3001 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3061 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3121 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3181 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3241 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3301 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3361 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3421 tgtacagcgc tggccaccac catcaccatc acggataccc atacgatgtt ccagattacg
     3481 ctagctgagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     3541 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3601 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3661 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3721 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3781 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3841 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3901 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3961 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4021 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4081 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4141 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4321 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4441 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4501 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4681 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4741 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4801 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4861 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4921 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4981 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5041 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5101 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5161 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5221 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5281 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5341 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5401 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5461 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5521 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5581 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5641 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     5701 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     5761 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5821 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5881 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5941 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     6001 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaattcct
     6061 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6121 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6181 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6241 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6301 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6361 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6421 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6481 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6541 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6601 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6661 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6721 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6781 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
     6841 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6901 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6961 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7021 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     7081 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     7141 tcctacttgg cagtacatct acgtattagt catcgctatt accatg
//
