LOCUS       sequence-245544-        4696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses peroxisomal roGFP2 (po-roGFP2) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4696)
  AUTHORS   Ivashchenko O, Van Veldhoven PP, Brees C, Ho YS, Terlecky SR, 
            Fransen M
  TITLE     Intraperoxisomal redox balance in mammalian cells: oxidative stress 
            and interorganellar cross-talk.
  JOURNAL   Mol Biol Cell. 2011 May;22(9):1440-51. doi: 10.1091/mbc.E10-11-0919.
            Epub 2011 Mar 3.
  PUBMED    21372177
REFERENCE   2  (bases 1 to 4696)
  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"; doi: 
            "10.1091/mbc.E10-11-0919"; journalName: "Mol Biol Cell"; date: 
            "2011-05"; volume: "22"; issue: "9"; pages: "1440-51"
COMMENT     Sequence Label: pMF1706
FEATURES             Location/Qualifiers
     source          1..4696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    364..485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(401..420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     455..474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(492..947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(629..650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     841..860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        974..1078
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1080..1437
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1080..1271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1103..1123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1288..1423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1472..2266
                     /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             1472..2266
                     /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(1526..1545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2136..2155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2454..2473)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2498..2545
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2874..3462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3363..3382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3637..3940
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3941..4144
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4095..4115
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(4207..4209,4210..4212,4213..4696,1..227)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4252..4273)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4513..4532)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
       61 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
      121 cctgctccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      181 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaag tccaagctgt
      241 gagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
      301 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
      361 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
      421 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
      481 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
      541 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      601 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
      661 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      721 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      781 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      841 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      901 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
      961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     1081 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1261 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     1381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     1441 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     1501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1621 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     1681 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     1801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1981 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     2101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     2161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     2281 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     2341 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     2401 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     2461 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2521 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     2581 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     2641 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     2701 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     2761 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2941 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3001 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3061 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3541 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     3601 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3661 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3721 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3781 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3841 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3901 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3961 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4021 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4081 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4141 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag
     4201 cttaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4261 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4321 acctacggca agctgaccct gaagttcatc tccaccaccg gcaagctgcc cgtgccctgg
     4381 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     4441 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4501 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4561 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4621 gggcacaagc tggagtacaa ctacaactgc cacaacgtct atatcatggc cgacaagcag
     4681 aagaacggca tcaagg
//
