LOCUS       sequence-11273-e        7130 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 7130)
  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 7130)
  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(DK0)-EGFP
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      13..22
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(19..21,22..24,25..738)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(19..21,22..24,25..735)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(64..85)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(325..344)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     672..693
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    861..982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(898..917)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     952..971
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(989..1444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1126..1147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1338..1357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1471..1575
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1577..1934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1577..1768
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1600..1620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1785..1920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1969..2763
                     /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             1969..2763
                     /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(2023..2042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2633..2652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2951..2970)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2995..3042
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3371..3959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3860..3879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4134..4437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4438..4641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4592..4612
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
       61 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      121 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      181 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      241 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      301 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      361 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      421 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      481 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      541 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      601 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      661 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      721 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
      781 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
      841 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
      901 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      961 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     1021 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1081 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1141 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1201 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1261 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1321 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1381 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1441 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1561 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     1621 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1681 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1741 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1801 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1861 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1921 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     1981 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2041 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2101 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     2161 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2221 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2281 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2341 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2401 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2461 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     2521 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2581 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2641 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2701 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2761 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     2941 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3001 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3061 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3121 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3181 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     3241 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3301 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3361 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3421 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3481 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3541 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3601 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3661 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3721 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3781 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3841 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3901 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3961 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4021 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     4081 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4141 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     4201 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     4261 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     4321 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     4381 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     4441 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     4501 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     4561 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     4621 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
     4681 cagatctcga gctcaagctt gcgcgcgcca tggcttctgg agccgattca aaaggtgatg
     4741 acctatcaac agccattctc aaacagaaga accgtcccaa tcggttaatt gttgatgaag
     4801 ccatcaatga ggacaacagt gtggtgtcct tgtcccagcc caagatggat gaattgcagt
     4861 tgttccgagg tgacacagtg ttgctgaaag gaaagaagag acgagaagct gtttgcatcg
     4921 tcctttctga tgatacttgt tctgatgaga agattcggat gaatagagtt gttcggaata
     4981 accttcgtgt acgcctaggg gatgtcatca gcatccagcc atgccctgat gtgaagtacg
     5041 gcaaacgtat ccatgtgctg cccattgatg acacagtgga aggcattact ggtaatctct
     5101 tcgaggtata ccttaagccg tacttcctgg aagcgtatcg acccatccgg aaaggagaca
     5161 tttttcttgt ccgtggtggg atgcgtgctg tggagttcaa agtggtggaa acagatccta
     5221 gcccttattg cattgttgct ccagacacag tgatccactg cgaaggggag cctatcaaac
     5281 gagaggatga ggaagagtcc ttgaatgaag tagggtatga tgacattggt ggctgcagga
     5341 agcagctagc tcagataaag gagatggtgg aactgcccct gagacatcct gccctcttta
     5401 aggcaattgg tgtgaagcct cctagaggaa tcctgcttta cggacctcct ggaacaggaa
     5461 agaccctgat tgctcgagct gtagcaaatg agactggagc cttcttcttc ttgatcaatg
     5521 gtcctgagat catgagcaaa ttggctggtg agtctgagag caaccttcgt aaagcctttg
     5581 aggaggctga gaagaatgct cctgccatca tcttcattga tcagctagat gccatcgctc
     5641 ccaaaagaga gaaaactcat ggcgaggtgg agcggcgcat tgtatcacag ttgttgaccc
     5701 tcatggatgg cctaaagcag agggcacatg tgattgttat ggcagcaacc aacagaccca
     5761 acagcattga cccagctcta cggcgatttg gtcgctttga cagggaggta gatattggaa
     5821 ttcctgatgc tacaggacgc ttagagattc ttcagatcca taccaagaac atgaagctgg
     5881 cagatgatgt ggacctggaa caggtagcca atgagactca cgggcatgtg ggtgctgact
     5941 tagcagccct gtgctcagag gctgctctgc aagccatccg caagaagatg gatctcattg
     6001 acctagagga tgagaccatt gatgccgagg tcatgaactc tctagcagtt actatggatg
     6061 acttccggtg ggccttgagc cagagtaacc catcagcact gcgggaaacc gtggtagagg
     6121 tgccacaggt aacctgggaa gacatcgggg gcctagagga tgtcaaacgt gagctacagg
     6181 agctggtcca gtatcctgtg gagcacccag acaaattcct gaagtttggc atgacacctt
     6241 ccaagggagt tctgttctat ggacctcctg gctgtgggaa aactttgttg gccaaagcca
     6301 ttgctaatga atgccaggcc aacttcatct ccatcaaggg tcctgagctg ctcaccatgt
     6361 ggtttgggga gtctgaggcc aatgtcagag aaatctttga caaggcccgc caagctgccc
     6421 cctgtgtgct attctttgat cagctggatt cgattgccaa ggctcgtgga ggtaacattg
     6481 gagatggtgg tggggctgct gaccgagtca tcaaccagat cctgacagaa atggatggca
     6541 tgtccactaa aaaaaatgtg ttcatcattg gcgctaccaa ccggcctgac atcattgatc
     6601 ctgccatcct cagacctggc cgtcttgatc agctcatcta catcccactt cctgatgaga
     6661 agtcccgtgt tgccatcctc aaggctaacc tgcgcaagtc cccagttgcc aaggatgtgg
     6721 acttggagtt cctggctaaa atgactaatg gcttctctgg agctgacctg acagagattt
     6781 gccagcgtgc ttgcaagctg gccatccgtg aatccatcga gagtgagatt aggcgagaac
     6841 gagagaggca gacaaaccca tcagccatgg aggtagaaga ggatgatcca gtgcctgaga
     6901 tccgtcgaga tcactttgaa gaagccatgc gctttgcgcg ccgttctgtc agtgacaatg
     6961 acattcggaa gtatgagatg tttgcccaga cccttcagca gagtcggggc tttggcagct
     7021 tcagattccc ttcagggaac cagggtggag ctggccccag tcagggcagt ggaggcggca
     7081 caggtggcag tgtatacaca gaagacaatg atgatgacct gtatggctcg
//
