LOCUS       sequence-461563-        8414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PEX3-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8414)
  AUTHORS   Kurikawa Y, Koyama-Honda I, Tamura N, Koike S, Mizushima N
  TITLE     Organelle landscape analysis using a multiparametric particle-based 
            method.
  JOURNAL   PLoS Biol. 2024 Sep 17;22(9):e3002777. doi: 
            10.1371/journal.pbio.3002777. eCollection 2024 Sep.
  PUBMED    39288101
REFERENCE   2  (bases 1 to 8414)
  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: "PLoS 
            Biol."; date: "2024-09-17"; volume: "22(9):e3002777. doi"; pages: " 
            10.1371/journal.pbio.3002777. eCollection 2024 Sep"
COMMENT     Sequence Label: pMXs-IP-PEX3-EGFP
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(3081..3083,3084..3086,3087..3800)
                     /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(3081..3083,3084..3086,3087..3797)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3809..3811,3812..3814,3815..4528)
                     /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(3809..3811,3812..3814,3815..4525)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    join(4559..5133,5134..5136,5137..5145)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4726..4743)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4953..4972
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5146..5748
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5645..5665
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5919..6388
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6518..6535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6689..7277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6769..6788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7441..8232,8233..8301))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8064..8083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8302..8406)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ctgccatgct gaggtctgta tggaattttc tgaaacgcca caaaaagaaa tgcatcttcc
     1981 tgggcacggt ccttggagga gtatatattc tggggaaata tggacagaag aaaatcagag
     2041 aaatacagga aagggaggct gcagaataca ttgcccaagc acgacgacaa tatcattttg
     2101 aaagtaacca gaggacttgc aatatgacag tgctgtccat gcttccaaca ctgagagagg
     2161 ccttaatgca gcaactgaat tccgagagcc tcacagctct gctaaaaaac aggccttcaa
     2221 acaagctaga aatatgggag gatctgaaga taataagttt cacaagaagt actgtggctg
     2281 tatacagtac ctgtatgctg gttgttcttt tgcgggtcca gttaaacata attggtggat
     2341 atatttacct ggataatgca gcagttggca aaaatggcac tacaattctt gctcccccag
     2401 atgtccaaca gcagtattta tcaagtattc agcacctact tggagatggc ctgacagaat
     2461 tgatcactgt cattaaacaa gctgtgcaga aggttttagg aagtgtttct cttaaacatt
     2521 ctttgtccct tttggacttg gagcaaaaac taaaagaaat cagaaatctc gttgagcagc
     2581 ataagtcttc ttcttggatt aataaagatg gatccaaacc tttattatgc cattatatga
     2641 tgccagatga agaaactcca ttagcagtgc aggcctgtgg actttctcct cgagacatta
     2701 ccactattaa acttctcaat gaaactagag acatgttgga aagcccagat tttagtacag
     2761 ttttgaatac ctgtttaaac cgaggtttta gtagacttct agacaatatg gctgagttct
     2821 ttcgacctac tgaacaggac ctgcaacatg gtaactctat gaatagtctt tccagtgtca
     2881 gcctgccttt agctaagata attccaatag taaacggaca gatccattca gtttgcagtg
     2941 aaacacctag tcattttgtt caggatctgt tgacaatgga gcaagtgaaa gactttgctg
     3001 ctaatgtgta tgaagctttt agtacccctc agcaactgga gaaaagatcc ggaggatcag
     3061 gagggaattc aggcggccgc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3121 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3181 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3241 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3301 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3361 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3421 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3481 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3541 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3601 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3661 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3721 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3781 tggacgagct gtacaagtaa gcggccgcat ggtgagcaag ggcgaggagc tgttcaccgg
     3841 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3901 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3961 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4021 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4081 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4141 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4201 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     4261 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     4321 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4381 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     4441 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4501 tctcggcatg gacgagctgt acaagtaagc ggccgctacg taaattccgc cccccccccc
     4561 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     4621 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     4681 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     4741 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     4801 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     4861 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     4921 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     4981 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     5041 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     5101 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt taccgagtac
     5161 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     5221 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     5281 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5341 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5401 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5461 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5521 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5581 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5641 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5701 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgacgataaa
     5761 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     5821 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     5881 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     5941 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     6001 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     6061 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     6121 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     6181 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     6241 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     6301 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     6361 ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt
     6421 ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttc
//
