LOCUS       sequence-245542-        4684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytosolic roGFP2 (c-roGFP2) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4684)
  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 4684)
  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: pMF1707
FEATURES             Location/Qualifiers
     source          1..4684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(267..269,270..272,273..986)
                     /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(312..333)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(573..592)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     920..941
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1108..1229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1145..1164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1199..1218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1236..1691)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1373..1394)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1585..1604
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1718..1822
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1824..2181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1824..2015
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1847..1867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2032..2167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2216..3010
                     /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             2216..3010
                     /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(2270..2289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2880..2899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3198..3217)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3242..3289
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3618..4206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4107..4126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4381..4684
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tccaccaccg
      421 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      481 tcagccgcta ccctgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaactgc cacaacgtct
      721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      841 gccccgtgct gctgcccgac aaccactacc tgagcacctg ctccgccctg agcaaagacc
      901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      961 tcggcatgga cgagctgtac aagtaagcgg ccgcgactct agatcataat cagccatacc
     1021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     1081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     1141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     1201 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     1261 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     1321 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     1381 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     1441 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     1501 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1561 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     1621 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1681 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1801 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     1861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2101 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     2161 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     2221 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2281 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2341 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     2401 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2461 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2521 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2581 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2641 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2701 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     2761 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2821 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2881 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2941 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3001 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3061 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3121 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3181 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3241 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     3301 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     3361 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     3421 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     3481 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3541 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3601 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3661 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3721 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3781 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3841 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3901 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3961 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4021 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4081 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4141 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4201 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4261 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     4321 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     4381 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     4441 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     4501 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     4561 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     4621 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     4681 catg
//
