LOCUS       sequence-404938-        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 (d)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2947..2956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2953..2955,2956..2958,2959..3672)
                     /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(2998..3019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3259..3278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3606..3627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3832..3851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3886..3905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3923..4378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4060..4081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4272..4291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4405..4509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4511..4868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4511..4702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4534..4554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4719..4854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4903..5697
                     /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             4903..5697
                     /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(4957..4976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5567..5586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5885..5904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5929..5976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6305..6893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6794..6813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7068..7371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7372..7562,1..13)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7526..7546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgat gtggcgacta
       61 cgtcgggccg ctgtggcctg tgaggtctgc cagtctttag tgaaacacag ctctggaata
      121 aaaggaagtt taccactaca aaaactacat ctggtttcac gaagcattta tcattcacat
      181 catcctacct taaagcttca acgaccccaa ttaaggacat cctttcagca gttctcttct
      241 ctgacaaacc ttcctttacg taaactgaaa ttctctccaa ttaaatatgg ctaccagcct
      301 cgcaggaatt tttggccagc aagattagct acgagactct taaaacttcg ctatctcata
      361 ctaggatcgg ctgttggggg tggctacaca gccaaaaaga cttttgatca gtggaaagat
      421 atgataccgg accttagtga atataaatgg attgtgcctg acattgtgtg ggaaattgat
      481 gagtatatcg attttgagaa aattagaaaa gcccttccta attcagaaga ccttgtaaag
      541 ttagcaccag actttgacaa gattgttgaa agccttagct tattgaagga cttttttacc
      601 tcaggttctc cggaagaaac ggcgtttaga gcaacagatc gtggatctga aagtgacaag
      661 cattttagaa aggtgtcaga caaagagaaa attgaccaac ttcaggaaga acttctgcac
      721 actcagttga agtatcagag aatcttggaa cgattagaaa aggagaacaa agaattgaga
      781 aaattagtat tgcagaaaga tgacaaaggc attcatcata gaaagcttaa gaaatctttg
      841 attgacatgt attctgaagt tcttgatgtt ctctctgatt atgatgccag ttataatacg
      901 caagatcatc tgccacgggt tgttgtggtt ggagatcaga gtgctggaaa gactagtgtg
      961 ttggaaatga ttgcccaagc tcgaatattc ccaagaggat ctggggagat gatgacacgt
     1021 tctccagtta aggtgactct gagtgaaggt cctcaccatg tggccctatt taaagatagt
     1081 tctcgggagt ttgatcttac caaagaagaa gatcttgcag cattaagaca tgaaatagaa
     1141 cttcgaatga ggaaaaatgt gaaagaaggc tgtaccgtta gccctgagac catatcctta
     1201 aatgtaaaag gccctggact acagaggatg gtgcttgttg acttaccagg tgtgattaat
     1261 actgtgacat caggcatggc tcctgacaca aaggaaacta ttttcagtat cagcaaagct
     1321 tacatgcaga atcctaatgc catcatactg tgtattcaag atggatctgt ggatgctgaa
     1381 cgcagtattg ttacagactt ggtcagtcaa atggaccctc atggaaggag aaccatattc
     1441 gttttgacca aagtagacct ggcagagaaa aatgtagcca gtccaagcag gattcagcag
     1501 ataattgaag gaaagctctt cccaatgaaa gctttaggtt attttgctgt tgtaacagga
     1561 aaagggaaca gctctgaaag cattgaagct ataagagaat atgaagaaga gttttttcag
     1621 aattcaaagc tcctaaagac aagcatgcta aaggcacacc aagtgactac aagaaattta
     1681 agccttgcag tatcagactg cttttggaaa atggtacgag agtctgttga acaacaggct
     1741 gatagtttca aagcaacacg ttttaacctt gaaactgaat ggaagaataa ctatcctcgc
     1801 ctgcgggaac ttgaccggaa tgaactattt gaaaaagcta aaaatgaaat ccttgatgaa
     1861 gttatcagtc tgagccaggt tacaccaaaa cattgggagg aaatccttca acaatctttg
     1921 tgggaaagag tatcaactca tgtgattgaa aacatctacc ttccagctgc gcagaccatg
     1981 aattcaggaa cttttaacac cacagtggat atcaagctta aacagtggac tgataaacaa
     2041 cttcctaata aagcagtaga ggttgcttgg gagaccctac aagcggaatt ttcccgcttt
     2101 atgacagaac cgaaagggaa agagcatgat gacatatttg ataaacttaa agaggccgtt
     2161 aaggaagaaa gtattaaacg acacaagtgg aatgactttg cggaggacag cttgagggtt
     2221 attcaacaca atgctttgga agaccgatcc atatctgata aacagcaatg ggatgcagct
     2281 atttatttta tggaagaggc tctgcaggct cgtctcaagg atactgaaaa tgcaattgaa
     2341 aacatggtgg gtccagactg gaaaaagagg tggttatact ggaagaatcg gacccaagaa
     2401 cagtgtgttc acaatgaaac caagaatgaa ttggagaaga tgttgaaatg taatgaggag
     2461 cacccagctt atcttgcaag tgatgaaata accacagtcc ggaagaacct tgaatcccga
     2521 ggagtagaag tagatccaag cttgattaag gatacttggc atcaagttta tagaagacat
     2581 tttttaaaaa cagctctaaa ccattgtaac ctttgtcgaa gaggttttta ttactaccaa
     2641 aggcattttg tagattctga gttggaatgc aatgatgtgg tcttgttttg gcgtatacag
     2701 cgcatgcttg ctatcaccgc aaatacttta aggcaacaac ttacaaatac tgaagttagg
     2761 cgattagaga aaaatgttaa agaggtattg gaagattttg ctgaagatgg tgagaagaag
     2821 attaaattgc ttactggtaa acgcgttcaa ctggcggaag acctcaagaa agttagagaa
     2881 attcaagaaa aacttgatgc tttcattgaa gctcttcatc aggagaaagg gctagcgcta
     2941 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3001 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3061 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3121 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3181 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3241 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3301 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3361 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3421 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3481 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3541 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     3601 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3661 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3721 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3781 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3901 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3961 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4021 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4081 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4141 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4201 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4261 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4321 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4381 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4441 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4501 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4681 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4741 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4801 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4861 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4921 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4981 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5041 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5101 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5161 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5221 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5281 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5341 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5401 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5461 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5521 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5581 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5641 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5701 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5761 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5821 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5881 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5941 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6001 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6061 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6121 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6181 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6241 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6961 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     7021 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     7081 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     7141 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     7201 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     7261 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     7321 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     7381 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     7441 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     7501 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     7561 ta
//
