LOCUS       sequence-404956-        7562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of GFP-tagged hOpa1 isoform 1 mutant 
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7562)
  AUTHORS   Nyenhuis SB, Wu X, Strub MP, Yim YI, Stanton AE, Baena V, Syed ZA, 
            Canagarajah B, Hammer JA, Hinshaw JE
  TITLE     OPA1 helical structures give perspective to mitochondrial 
            dysfunction.
  JOURNAL   Nature. 2023 Aug 23. doi: 10.1038/s41586-023-06462-1.
  PUBMED    37612506
REFERENCE   2  (bases 1 to 7562)
  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: "Nature. 
            2023 Aug 23. doi: 10.1038/s41586-023-06462-1."
COMMENT     Sequence Label: Control-3
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..104
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3067..3076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3073..3075,3076..3078,3079..3792)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3118..3139)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3379..3398)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3726..3747
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3915..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3952..3971)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4006..4025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4043..4498)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4180..4201)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4392..4411
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4525..4629
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4631..4988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4631..4822
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4654..4674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4839..4974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5023..5817
                     /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             5023..5817
                     /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(5077..5096)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5687..5706
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6005..6024)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6049..6096
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6425..7013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6914..6933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7188..7491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7492..7562,1..133)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
       61 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      121 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgat gtggcgacta
      181 cgtcgggccg ctgtggcctg tgaggtctgc cagtctttag tgaaacacag ctctggaata
      241 aaaggaagtt taccactaca aaaactacat ctggtttcac gaagcattta tcattcacat
      301 catcctacct taaagcttca acgaccccaa ttaaggacat cctttcagca gttctcttct
      361 ctgacaaacc ttcctttacg taaactgaaa ttctctccaa ttaaatatgg ctaccagcct
      421 cgcaggaatt tttggccagc aagattagct acgagactct taaaacttcg ctatctcata
      481 ctaggatcgg ctgttggggg tggctacaca gccaaaaaga cttttgatca gtggaaagat
      541 atgataccgg accttagtga atataaatgg attgtgcctg acattgtgtg ggaaattgat
      601 gagtatatcg attttgagaa aattagaaaa gcccttccta attcagaaga ccttgtaaag
      661 ttagcaccag actttgacaa gattgttgaa agccttagct tattgaagga cttttttacc
      721 tcaggttctc cggaagaaac ggcgtttaga gcaacagatc gtggatctga aagtgacaag
      781 cattttagaa aggtgtcaga caaagagaaa attgaccaac ttcaggaaga acttctgcac
      841 actcagttga agtatcagag aatcttggaa cgattagaaa aggagaacaa agaattgaga
      901 aaattagtat tgcagaaaga tgacaaaggc attcatcata gaaagcttaa gaaatctttg
      961 attgacatgt attctgaagt tcttgatgtt ctctctgatt atgatgccag ttataatacg
     1021 caagatcatc tgccacgggt tgttgtggtt ggagatcaga gtgctggaaa gactagtgtg
     1081 ttggaaatga ttgcccaagc tcgaatattc ccaagaggat ctggggagat gatgacacgt
     1141 tctccagtta aggtgactct gagtgaaggt cctcaccatg tggccctatt taaagatagt
     1201 tctcgggagt ttgatcttac caaagaagaa gatcttgcag cattaagaca tgaaatagaa
     1261 cttcgaatga gggccaatgt gaaagaaggc tgtaccgtta gccctgagac catatcctta
     1321 aatgtaaaag gccctggact acagaggatg gtgcttgttg acttaccagg tgtgattaat
     1381 actgtgacat caggcatggc tcctgacaca aaggaaacta ttttcagtat cagcaaagct
     1441 tacatgcaga atcctaatgc catcatactg tgtattcaag atggatctgt ggatgctgaa
     1501 cgcagtattg ttacagactt ggtcagtcaa atggaccctc atggaaggag aaccatattc
     1561 gttttgacca aagtagacct ggcagagaaa aatgtagcca gtccaagcag gattcagcag
     1621 ataattgaag gaaagctctt cccaatgaaa gctttaggtt attttgctgt tgtaacagga
     1681 aaagggaaca gctctgaaag cattgaagct ataagagaat atgaagaaga gttttttcag
     1741 aattcaaagc tcctaaagac aagcatgcta aaggcacacc aagtgactac aagaaattta
     1801 agccttgcag tatcagactg cttttggaaa atggtacgag agtctgttga acaacaggct
     1861 gatagtttca aagcaacacg ttttaacctt gaaactgaat ggaagaataa ctatcctcgc
     1921 ctgcgggaac ttgaccggaa tgaactattt gaaaaagcta aaaatgaaat ccttgatgaa
     1981 gttatcagtc tgagccaggt tacaccaaaa cattgggagg aaatccttca acaatctttg
     2041 tgggaaagag tatcaactca tgtgattgaa aacatctacc ttccagctgc gcagaccatg
     2101 aattcaggaa cttttaacac cacagtggat atcaagctta aacagtggac tgataaacaa
     2161 cttcctaata aagcagtaga ggttgcttgg gagaccctac aagaagaatt ttcccgcttt
     2221 atgacagaac cgaaagggaa agagcatgat gacatatttg ataaacttaa agaggccgtt
     2281 aaggaagaaa gtattaaacg acacaagtgg aatgactttg cggaggacag cttgagggtt
     2341 attcaacaca atgctttgga agaccgatcc atatctgata aacagcaatg ggatgcagct
     2401 atttatttta tggaagaggc tctgcaggct cgtctcaagg atactgaaaa tgcaattgaa
     2461 aacatggtgg gtccagactg gaaaaagagg tggttatact ggaagaatcg gacccaagaa
     2521 cagtgtgttc acaatgaaac caagaatgaa ttggagaaga tgttgaaatg taatgaggag
     2581 cacccagctt atcttgcaag tgatgaaata accacagtcc ggaagaacct tgaatcccga
     2641 ggagtagaag tagatccaag cttgattaag gatacttggc atcaagttta tagaagacat
     2701 tttttaaaaa cagctctaaa ccattgtaac ctttgtcgaa gaggttttta ttactaccaa
     2761 aggcattttg tagattctga gttggaatgc aatgatgtgg tcttgttttg gcgtatacag
     2821 cgcatgcttg ctatcaccgc aaatacttta aggcaacaac ttacaaatac tgaagttagg
     2881 cgattagaga aaaatgttaa agaggtattg gaagattttg ctgaagatgg tgagaagaag
     2941 attaaattgc ttactggtaa acgcgttcaa ctggcggaag acctcaagaa agttagagaa
     3001 attcaagaaa aacttgatgc tttcattgaa gctcttcatc aggagaaagg gctagcgcta
     3061 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3121 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3181 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3241 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3301 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3361 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3421 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3481 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3541 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3601 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3661 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     3721 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3781 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3841 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3901 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3961 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4021 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     4081 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4141 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4201 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4261 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4321 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4381 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4441 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4621 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4981 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5041 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5101 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5161 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5221 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5281 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5341 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5401 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5461 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5521 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5581 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5641 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5701 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5761 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5821 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5881 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5941 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     6001 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     6061 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6121 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6181 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6241 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     7141 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     7201 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     7261 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     7321 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     7381 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     7441 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     7501 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     7561 ca
//
