LOCUS       sequence-404937-        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: Interface-1 (c)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2967..2976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2973..2975,2976..2978,2979..3692)
                     /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(3018..3039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3279..3298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3626..3647
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3815..3936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3852..3871)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3906..3925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3943..4398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4080..4101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4292..4311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4425..4529
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4531..4888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4531..4722
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4554..4574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4739..4874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4923..5717
                     /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             4923..5717
                     /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(4977..4996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5587..5606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5905..5924)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5949..5996
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6325..6913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6814..6833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7088..7391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7392..7562,1..33)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7546..7562,1..4)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
       61 cgctaccgat gtggcgacta cgtcgggccg ctgtggcctg tgaggtctgc cagtctttag
      121 tgaaacacag ctctggaata aaaggaagtt taccactaca aaaactacat ctggtttcac
      181 gaagcattta tcattcacat catcctacct taaagcttca acgaccccaa ttaaggacat
      241 cctttcagca gttctcttct ctgacaaacc ttcctttacg taaactgaaa ttctctccaa
      301 ttaaatatgg ctaccagcct cgcaggaatt tttggccagc aagattagct acgagactct
      361 taaaacttcg ctatctcata ctaggatcgg ctgttggggg tggctacaca gccaaaaaga
      421 cttttgatca gtggaaagat atgataccgg accttagtga atataaatgg attgtgcctg
      481 acattgtgtg ggaaattgat gagtatatcg attttgagaa aattagaaaa gcccttccta
      541 attcagaaga ccttgtaaag ttagcaccag actttgacaa gattgttgaa agccttagct
      601 tattgaagga cttttttacc tcaggttctc cggaagaaac ggcgtttaga gcaacagatc
      661 gtggatctga aagtgacaag cattttagaa aggtgtcaga caaagagaaa attgaccaac
      721 ttcaggaaga acttctgcac actcagttga agtatcagag aatcttggaa cgattagaaa
      781 aggagaacaa agaattgaga aaattagtat tgcagaaaga tgacaaaggc attcatcata
      841 gaaagcttaa gaaatctttg attgacatgt attctgaagt tcttgatgtt ctctctgatt
      901 atgatgccag ttataatacg caagatcatc tgccacgggt tgttgtggtt ggagatcaga
      961 gtgctggaaa gactagtgtg ttggaaatga ttgcccaagc tcgaatattc ccaagaggat
     1021 ctggggagat gatgacacgt tctccagtta aggtgactct gagtgaaggt cctcaccatg
     1081 tggccctatt taaagatagt tctcgggagt ttgatcttac caaagaagaa gatcttgcag
     1141 cattaagaca tgaaatagaa cttcgaatga ggaaaaatgt gaaagaaggc tgtaccgtta
     1201 gccctgagac catatcctta aatgtaaaag gccctggact acagaggatg gtgcttgttg
     1261 acttaccagg tgtgattaat actgtgacat caggcatggc tcctgacaca aaggaaacta
     1321 ttttcagtat cagcaaagct tacatgcaga atcctaatgc catcatactg tgtattcaag
     1381 atggatctgt ggatgctgaa cgcagtattg ttacagactt ggtcagtcaa atggaccctc
     1441 atggaaggag aaccatattc gttttgacca aagtagacct ggcagagaaa aatgtagcca
     1501 gtccaagcag gattcagcag ataattgaag gaaagctctt cccaatgaaa gctttaggtt
     1561 attttgctgt tgtaacagga aaagggaaca gctctgaaag cattgaagct ataagagaat
     1621 atgaagaaga gttttttcag aattcaaagc tcctaaagac aagcatgcta aaggcacacc
     1681 aagtgactac aagaaattta agccttgcag tatcagactg cttttggaaa atggtacgag
     1741 agtctgttga acaacaggct gatagtttca aagcaacacg ttttaacctt gaaactgaat
     1801 ggaagaataa ctatcctcgc ctgcgggaac ttgaccggaa tgaactattt gaaaaagcta
     1861 aaaatgaaat ccttgatgaa gttatcagtc tgagccaggt tacaccaaaa cattgggagg
     1921 aaatccttca acaatctttg tgggaaagag tatcaactca tgtgattgaa aacatctacc
     1981 ttccagctgc gcagaccatg aattcaggaa cttttaacac cacagtggat atcaagctta
     2041 aagcgtggac tgataaacaa cttcctaata aagcagtaga ggttgcttgg gagaccctac
     2101 aagaagaatt ttcccgcttt atgacagaac cgaaagggaa agagcatgat gacatatttg
     2161 ataaacttaa agaggccgtt aaggaagaaa gtattaaacg acacaagtgg aatgactttg
     2221 cggaggacag cttgagggtt attcaacaca atgctttgga agaccgatcc atatctgata
     2281 aacagcaatg ggatgcagct atttatttta tggaagaggc tctgcaggct cgtctcaagg
     2341 atactgaaaa tgcaattgaa aacatggtgg gtccagactg gaaaaagagg tggttatact
     2401 ggaagaatcg gacccaagaa cagtgtgttc acaatgaaac caagaatgaa ttggagaaga
     2461 tgttgaaatg taatgaggag cacccagctt atcttgcaag tgatgaaata accacagtcc
     2521 ggaagaacct tgaatcccga ggagtagaag tagatccaag cttgattaag gatacttggc
     2581 atcaagttta tagaagacat tttttaaaaa cagctctaaa ccattgtaac ctttgtcgaa
     2641 gaggttttta ttactaccaa aggcattttg tagattctga gttggaatgc aatgatgtgg
     2701 tcttgttttg gcgtatacag cgcatgcttg ctatcaccgc aaatacttta aggcaacaac
     2761 ttacaaatac tgaagttagg cgattagaga aaaatgttaa agaggtattg gaagattttg
     2821 ctgaagatgg tgagaagaag attaaattgc ttactggtaa acgcgttcaa ctggcggaag
     2881 acctcaagaa agttagagaa attcaagaaa aacttgatgc tttcattgaa gctcttcatc
     2941 aggagaaagg gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     3001 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3061 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3121 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     3181 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3241 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3301 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3361 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3421 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3481 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3541 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     3601 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3661 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     3721 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3781 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3901 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3961 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4021 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4081 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4141 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4201 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4261 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4321 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4381 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4501 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4561 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4681 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4741 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4801 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4861 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4921 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5101 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5761 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5881 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5941 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6001 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6061 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6121 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6961 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7021 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     7081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     7141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     7201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     7261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     7321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     7381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     7441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     7501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     7561 gg
//
