LOCUS       sequence-323838-        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(R155H)-EGFP
FEATURES             Location/Qualifiers
     source          1..7186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3061..3070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3067..3069,3070..3072,3073..3780)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(3067..3069,3070..3072,3073..3780)
                     /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(3112..3133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3373..3392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3720..3741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3790..3807
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3811..3837
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3965..4086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4002..4021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4056..4075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4093..4548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4230..4251)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4442..4461
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4575..4679
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4681..5038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4681..4872
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4704..4724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4889..5024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5073..5867
                     /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             5073..5867
                     /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(5127..5146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5737..5756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6055..6074)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6099..6146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6475..7063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6964..6983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatctcgagc tcaagcttgc gcgcgccatg gcttctggag ccgattcaaa aggtgatgac
      661 ctatcaacag ccattctcaa acagaagaac cgtcccaatc ggttaattgt tgatgaagcc
      721 atcaatgagg acaacagtgt ggtgtccttg tcccagccca agatggatga attgcagttg
      781 ttccgaggtg acacagtgtt gctgaaagga aagaagagac gagaagctgt ttgcatcgtc
      841 ctttctgatg atacttgttc tgatgagaag attcggatga atagagttgt tcggaataac
      901 cttcgtgtac gcctagggga tgtcatcagc atccagccat gccctgatgt gaagtacggc
      961 aaacgtatcc atgtgctgcc cattgatgac acagtggaag gcattactgg taatctcttc
     1021 gaggtatacc ttaagccgta cttcctggaa gcgtatcgac ccatccggaa aggagacatt
     1081 tttcttgtcc atggtgggat gcgtgctgtg gagttcaaag tggtggaaac agatcctagc
     1141 ccttattgca ttgttgctcc agacacagtg atccactgcg aaggggagcc tatcaaacga
     1201 gaggatgagg aagagtcctt gaatgaagta gggtatgatg acattggtgg ctgcaggaag
     1261 cagctagctc agataaagga gatggtggaa ctgcccctga gacatcctgc cctctttaag
     1321 gcaattggtg tgaagcctcc tagaggaatc ctgctttacg gacctcctgg aacaggaaag
     1381 accctgattg ctcgagctgt agcaaatgag actggagcct tcttcttctt gatcaatggt
     1441 cctgagatca tgagcaaatt ggctggtgag tctgagagca accttcgtaa agcctttgag
     1501 gaggctgaga agaatgctcc tgccatcatc ttcattgatg agctagatgc catcgctccc
     1561 aaaagagaga aaactcatgg cgaggtggag cggcgcattg tatcacagtt gttgaccctc
     1621 atggatggcc taaagcagag ggcacatgtg attgttatgg cagcaaccaa cagacccaac
     1681 agcattgacc cagctctacg gcgatttggt cgctttgaca gggaggtaga tattggaatt
     1741 cctgatgcta caggacgctt agagattctt cagatccata ccaagaacat gaagctggca
     1801 gatgatgtgg acctggaaca ggtagccaat gagactcacg ggcatgtggg tgctgactta
     1861 gcagccctgt gctcagaggc tgctctgcaa gccatccgca agaagatgga tctcattgac
     1921 ctagaggatg agaccattga tgccgaggtc atgaactctc tagcagttac tatggatgac
     1981 ttccggtggg ccttgagcca gagtaaccca tcagcactgc gggaaaccgt ggtagaggtg
     2041 ccacaggtaa cctgggaaga catcgggggc ctagaggatg tcaaacgtga gctacaggag
     2101 ctggtccagt atcctgtgga gcacccagac aaattcctga agtttggcat gacaccttcc
     2161 aagggagttc tgttctatgg acctcctggc tgtgggaaaa ctttgttggc caaagccatt
     2221 gctaatgaat gccaggccaa cttcatctcc atcaagggtc ctgagctgct caccatgtgg
     2281 tttggggagt ctgaggccaa tgtcagagaa atctttgaca aggcccgcca agctgccccc
     2341 tgtgtgctat tctttgatga gctggattcg attgccaagg ctcgtggagg taacattgga
     2401 gatggtggtg gggctgctga ccgagtcatc aaccagatcc tgacagaaat ggatggcatg
     2461 tccactaaaa aaaatgtgtt catcattggc gctaccaacc ggcctgacat cattgatcct
     2521 gccatcctca gacctggccg tcttgatcag ctcatctaca tcccacttcc tgatgagaag
     2581 tcccgtgttg ccatcctcaa ggctaacctg cgcaagtccc cagttgccaa ggatgtggac
     2641 ttggagttcc tggctaaaat gactaatggc ttctctggag ctgacctgac agagatttgc
     2701 cagcgtgctt gcaagctggc catccgtgaa tccatcgaga gtgagattag gcgagaacga
     2761 gagaggcaga caaacccatc agccatggag gtagaagagg atgatccagt gcctgagatc
     2821 cgtcgagatc actttgaaga agccatgcgc tttgcgcgcc gttctgtcag tgacaatgac
     2881 attcggaagt atgagatgtt tgcccagacc cttcagcaga gtcggggctt tggcagcttc
     2941 agattccctt cagggaacca gggtggagct ggccccagtc agggcagtgg aggcggcaca
     3001 ggtggcagtg tatacacaga agacaatgat gatgacctgt atggctcgga tccaccggtc
     3061 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     3121 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     3181 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     3241 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     3301 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     3361 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     3421 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3481 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     3541 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     3601 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3661 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     3721 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     3781 agcgctggcc accaccatca ccatcacgga tacccatacg atgttccaga ttacgctagc
     3841 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3901 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3961 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4081 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4141 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4201 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4261 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4321 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4381 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4441 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4501 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4681 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4921 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4981 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5041 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5101 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5161 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5221 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5281 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5341 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5401 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5461 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5521 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5581 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5641 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5701 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5761 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5821 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5881 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5941 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6001 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6061 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6121 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6181 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6241 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6301 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6361 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7141 ccctgattct gtggataacc gtattaccgc catgcattag ttatta
//
