LOCUS       sequence-403805-        7392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged MVP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  AUTHORS   Kurusu R, Fujimoto Y, Morishita H, Noshiro D, Takada S, Yamano K, 
            Tanaka H, Arai R, Kageyama S, Funakoshi T, Komatsu-Hirota S, Taka H,
            Kazuno S, Miura Y, Koike M, Wakai T, Waguri S, Noda NN, Komatsu M
  TITLE     Integrated proteomics identifies p62-dependent selective autophagy 
            of the supramolecular vault complex.
  JOURNAL   Dev Cell. 2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015.
  PUBMED    37192622
REFERENCE   2  (bases 1 to 7392)
  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: "Dev Cell. 
            2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015."
COMMENT     Sequence Label: pEGFP-C2-MVP
FEATURES             Location/Qualifiers
     source          1..7392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4180..4301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4217..4236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4271..4290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4308..4763)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4445..4466)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4657..4676
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4790..4894
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4896..5253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4896..5087
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4919..4939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5104..5239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5288..6082
                     /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             5288..6082
                     /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(5342..5361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5952..5971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6270..6289)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6314..6361
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6690..7278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7179..7198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcgaattcat ggcaactgaa
     1381 gagttcatca tccgcatccc cccataccac tatatccatg tgctggacca gaacagcaac
     1441 gtgtcccgtg tggaggtcgg gccaaagacc tacatccggc aggacaatga gagggtactg
     1501 tttgccccca tgcgcatggt gaccgtcccc ccacgtcact actgcacagt ggccaaccct
     1561 gtgtctcggg atgcccaggg cttggtgctg tttgatgtca cagggcaagt tcggcttcgc
     1621 cacgctgacc tcgagatccg gctggcccag gaccccttcc ccctgtaccc aggggaggtg
     1681 ctggaaaagg acatcacacc cctgcaggtg gttctgccca acactgccct ccatctaaag
     1741 gcgctgcttg attttgagga taaagatgga gacaaggtgg tggcaggaga tgagtggctt
     1801 ttcgagggac ctggcacgta catcccccgg aaggaagtgg aggtcgtgga gatcattcag
     1861 gccaccatca tcaggcagaa ccaggctctg cggctcaggg cccgcaagga gtgctgggac
     1921 cgggacggca aggagagggt gacaggggaa gaatggctgg tcaccacagt aggggcgtac
     1981 ctcccagcgg tgtttgagga ggttctggat ttggtggacg ccgtcatcct tacggaaaag
     2041 acagccctgc acctccgggc tcggcggaac ttccgggact tcaggggagt gtcccgccgc
     2101 actggggagg agtggctggt aacagtgcag gacacagagg cccacgtgcc agatgtccac
     2161 gaggaggtgc tgggggttgt gcccatcacc accctgggcc cccacaacta ctgcgtgatt
     2221 ctcgaccctg tcggaccgga tggcaagaat cagctggggc agaagcgcgt ggtcaaggga
     2281 gagaagtctt ttttcctcca gccaggagag cagctggaac aaggcatcca ggatgtgtat
     2341 gtgctgtcgg agcagcaggg gctgctgctg agggccctgc agcccctgga ggagggggag
     2401 gatgaggaga aggtctcaca ccaggctggg gaccactggc tcatccgcgg acccctggag
     2461 tatgtgccat ctgccaaagt ggaggtggtg gaggagcgcc aggccatccc tctagacgag
     2521 aacgagggca tctatgtgca ggatgtcaag accggaaagg tgcgcgctgt gattggaagc
     2581 acctacatgc tgacccagga cgaagtcctg tgggagaaag agctgcctcc cggggtggag
     2641 gagctgctga acaaggggca ggaccctctg gcagacaggg gtgagaagga cacagctaag
     2701 agcctccagc ccttggcgcc ccggaacaag acccgtgtgg tcagctaccg cgtgccccac
     2761 aacgctgcgg tgcaggtgta cgactaccga gagaagcgag cccgcgtggt cttcgggcct
     2821 gagctggtgt cgctgggtcc tgaggagcag ttcacagtgt tgtccctctc agctgggcgg
     2881 cccaagcgtc cccatgcccg ccgtgcgctc tgcctgctgc tggggcctga cttcttcaca
     2941 gacgtcatca ccatcgaaac ggcggatcat gccaggctgc aactgcagct ggcctacaac
     3001 tggcactttg aggtgaatga ccggaaggac ccccaagaga cggccaagct cttttcagtg
     3061 ccagactttg taggtgatgc ctgcaaagcc atcgcatccc gggtgcgggg ggccgtggcc
     3121 tctgtcactt tcgatgactt ccataagaac tcagcccgca tcattcgcac tgctgtcttt
     3181 ggctttgaga cctcggaagc gaagggcccc gatggcatgg ccctgcccag gccccgggac
     3241 caggctgtct tcccccaaaa cgggctggtg gtcagcagtg tggacgtgca gtcagtggag
     3301 cctgtggatc agaggacccg ggacgccctg caacgcagcg tccagctggc catcgagatc
     3361 accaccaact cccaggaagc ggcggccaag catgaggctc agagactgga gcaggaagcc
     3421 cgcggccggc ttgagcggca gaagatcctg gaccagtcag aagccgagaa agctcgcaag
     3481 gaacttttgg agctggaggc tctgagcatg gccgtggaga gcaccgggac tgccaaggcg
     3541 gaggccgagt cccgtgcgga ggcagcccgg attgagggag aagggtccgt gctgcaggcc
     3601 aagctaaaag cacaggcctt ggccattgaa acggaggctg agctccagag ggtccagaag
     3661 gtccgagagc tggaactggt ctatgcccgg gcccagctgg agctggaggt gagcaaggct
     3721 cagcagctgg ctgaggtgga ggtgaagaag ttcaagcaga tgacagaggc cataggcccc
     3781 agcaccatca gggaccttgc tgtggctggg cctgagatgc aggtaaaact gctccagtcc
     3841 ctgggcctga aatcaaccct catcaccgat ggctccactc ccatcaacct cttcaacaca
     3901 gcctttgggc tgctggggat ggggcccgag ggtcagcccc tgggcagaag ggtggccagt
     3961 gggcccagcc ctggggaggg gatatccccc cagtctgctc aggcccctca agctcctgga
     4021 gacaaccacg tggtgcctgt actgcgctaa agatccaccg gatctagata actgatcata
     4081 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4141 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4201 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4261 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     4321 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4381 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4441 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4501 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4561 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4621 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4681 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4741 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4801 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4861 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4921 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4981 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5041 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5101 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5161 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5221 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5281 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5341 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5401 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5461 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5521 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5581 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5641 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5701 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5761 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5821 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5881 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5941 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6001 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6061 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6121 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6181 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6241 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6301 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6361 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6421 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6481 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     6541 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6601 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6661 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6721 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6781 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6841 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6901 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6961 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7021 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7081 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7141 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7201 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7261 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7321 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7381 accgccatgc at
//
