LOCUS       sequence-416289-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitophagy marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  AUTHORS   Nakamura H, Rho E, Lee CT, Itoh K, Deng D, Watanabe S, Razavi S, 
            Matsubayashi HT, Zhu C, Jung E, Rangamani P, Watanabe S, Inoue T
  TITLE     ActuAtor, a Listeria-inspired molecular tool for physical 
            manipulation of intracellular organizations through de novo actin 
            polymerization.
  JOURNAL   Cell Rep. 2023 Sep 2:113089. doi: 10.1016/j.celrep.2023.113089.
  PUBMED    37734382
REFERENCE   2  (bases 1 to 6096)
  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: "Cell Rep. 
            2023 Sep 2:113089. doi: 10.1016/j.celrep.2023.113089."
COMMENT     Sequence Label: EYFP-Parkin
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     135..155
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      223..232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(229..231,232..234,235..945)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(274..295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(535..554)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     882..903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2500..2621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2537..2556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2591..2610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2628..3083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2765..2786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2977..2996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3110..3214
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3216..3573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3216..3407
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3239..3259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3424..3559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3608..4402
                     /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             3608..4402
                     /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(3662..3681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4272..4291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4590..4609)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4634..4681
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5010..5598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5499..5518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5773..6076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6077..6096,1..184)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
       61 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      121 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      181 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag
      241 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      301 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      361 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      421 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      481 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      541 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      601 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      661 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
      721 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      781 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
      841 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      901 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
      961 cgagctcaag ctttgatagt gtttgtcagg ttcaactcca gccatggttt cccagtggag
     1021 gtcgattctg acaccagcat cttccagctc aaggaggtgg ttgctaagcg acagggggtt
     1081 ccggctgacc agttgcgtgt gattttcgca gggaaggagc tgaggaatga ctggactgtg
     1141 cagaattgtg acctggatca gcagagcatt gttcacattg tgcagagacc gtggagaaaa
     1201 ggtcaagaaa tgaatgcaac tggaggcgac gaccccagaa acgcggcggg aggctgtgag
     1261 cgggagcccc agagcttgac tcgggtggac ctcagcagct cagtcctccc aggagactct
     1321 gtggggctgg ctgtcattct gcacactgac agcaggaagg actcaccacc agctggaagt
     1381 ccagcaggta gatcaatcta caacagcttt tatgtgtatt gcaaaggccc ctgtcaaaga
     1441 gtgcagccgg gaaaactcag ggtacagtgc agcacctgca ggcaggcaac gctcaccttg
     1501 acccagggtc catcttgctg ggatgatgtt ttaattccaa accggatgag tggtgaatgc
     1561 caatccccac actgccctgg gactagtgca gaatttttct ttaaatgtgg agcacacccc
     1621 acctctgaca aggaaacatc agtagctttg cacctgatcg caacaaatag tcggaacatc
     1681 acttgcatta cgtgcacaga cgtcaggagc cccgtcctgg ttttccagtg caactcccgc
     1741 cacgtgattt gcttagactg tttccactta tactgtgtga caagactcaa tgatcggcag
     1801 tttgtccacg accctcagct tggctactcc ctgccttgtg tggctggctg tcccaactcc
     1861 ttgattaaag agctccatca cttcaggatt ctgggagaag agcagtacaa ccggtaccag
     1921 cagtatggtg cagaggagtg tgtcctgcag atggggggcg tgttatgccc ccgccctggc
     1981 tgtggagcgg ggctgctgcc ggagcctgac cagaggaaag tcacctgcga agggggcaat
     2041 ggcctgggct gtgggtttgc cttctgccgg gaatgtaaag aagcgtacca tgaaggggag
     2101 tgcagtgccg tatttgaagc ctcaggaaca actactcagg cctacagagt cgatgaaaga
     2161 gccgccgagc aggctcgttg ggaagcagcc tccaaagaaa ccatcaagaa aaccaccaag
     2221 ccctgtcccc gctgccatgt accagtggaa aaaaatggag gctgcatgca catgaagtgt
     2281 ccgcagcccc agtgcaggct cgagtggtgc tggaactgtg gctgcgagtg gaaccgcgtc
     2341 tgcatggggg accactggtt cgacgtgtaa ggatccaccg gatctagata actgatcata
     2401 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2461 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2581 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2641 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2701 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2761 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2821 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2881 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2941 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3001 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3061 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3181 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3241 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3301 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3421 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3481 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3541 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3601 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3661 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3721 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3781 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3841 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3901 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3961 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4021 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4081 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4141 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4201 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4261 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4321 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4381 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4441 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4501 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4561 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4621 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4681 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4741 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4801 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4861 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4921 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5641 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5701 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5761 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5821 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5881 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5941 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     6001 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     6061 catcgctatt accatggtga tgcggttttg gcagta
//
