LOCUS       sequence-403658-        7341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-tagged MVP delta607-623aa in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  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 7341)
  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 delta607-623aa
FEATURES             Location/Qualifiers
     source          1..7341
                     /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    4129..4250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4166..4185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4220..4239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4257..4712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4394..4415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4606..4625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4739..4843
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4845..5202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4845..5036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4868..4888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5053..5188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5237..6031
                     /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             5237..6031
                     /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(5291..5310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5901..5920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6219..6238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6263..6310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6639..7227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7128..7147
                     /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 ggctttgacc aggctgtctt cccccaaaac gggctggtgg tcagcagtgt ggacgtgcag
     3241 tcagtggagc ctgtggatca gaggacccgg gacgccctgc aacgcagcgt ccagctggcc
     3301 atcgagatca ccaccaactc ccaggaagcg gcggccaagc atgaggctca gagactggag
     3361 caggaagccc gcggccggct tgagcggcag aagatcctgg accagtcaga agccgagaaa
     3421 gctcgcaagg aacttttgga gctggaggct ctgagcatgg ccgtggagag caccgggact
     3481 gccaaggcgg aggccgagtc ccgtgcggag gcagcccgga ttgagggaga agggtccgtg
     3541 ctgcaggcca agctaaaagc acaggccttg gccattgaaa cggaggctga gctccagagg
     3601 gtccagaagg tccgagagct ggaactggtc tatgcccggg cccagctgga gctggaggtg
     3661 agcaaggctc agcagctggc tgaggtggag gtgaagaagt tcaagcagat gacagaggcc
     3721 ataggcccca gcaccatcag ggaccttgct gtggctgggc ctgagatgca ggtaaaactg
     3781 ctccagtccc tgggcctgaa atcaaccctc atcaccgatg gctccactcc catcaacctc
     3841 ttcaacacag cctttgggct gctggggatg gggcccgagg gtcagcccct gggcagaagg
     3901 gtggccagtg ggcccagccc tggggagggg atatcccccc agtctgctca ggcccctcaa
     3961 gctcctggag acaaccacgt ggtgcctgta ctgcgctaaa gatccaccgg atctagataa
     4021 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4081 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4141 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4201 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     4261 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4321 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4381 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4441 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4501 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4561 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4621 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4681 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4741 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4801 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4861 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4921 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5041 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5101 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5161 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5221 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5281 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5341 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5401 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5461 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5521 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5581 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5641 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5701 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5761 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5821 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5881 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5941 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6001 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6061 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6121 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6181 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6241 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6301 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6361 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6421 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6481 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6781 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7321 aaccgtatta ccgccatgca t
//
