LOCUS       sequence-410673-        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-4 (b)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2975..2984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2981..2983,2984..2986,2987..3700)
                     /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(3026..3047)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3287..3306)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3634..3655
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3823..3944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3860..3879)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3914..3933
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3951..4406)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4088..4109)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4300..4319
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4539..4896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4539..4730
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4562..4582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4747..4882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4931..5725
                     /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             4931..5725
                     /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(4985..5004)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5595..5614
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5913..5932)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5957..6004
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6333..6921
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6822..6841
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7096..7399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7400..7562,1..41)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7554..7562,1..12)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
       61 tccgctagcg ctaccgatgt ggcgactacg tcgggccgct gtggcctgtg aggtctgcca
      121 gtctttagtg aaacacagct ctggaataaa aggaagttta ccactacaaa aactacatct
      181 ggtttcacga agcatttatc attcacatca tcctacctta aagcttcaac gaccccaatt
      241 aaggacatcc tttcagcagt tctcttctct gacaaacctt cctttacgta aactgaaatt
      301 ctctccaatt aaatatggct accagcctcg caggaatttt tggccagcaa gattagctac
      361 gagactctta aaacttcgct atctcatact aggatcggct gttgggggtg gctacacagc
      421 caaaaagact tttgatcagt ggaaagatat gataccggac cttagtgaat ataaatggat
      481 tgtgcctgac attgtgtggg aaattgatga gtatatcgat tttgagaaaa ttagaaaagc
      541 ccttcctaat tcagaagacc ttgtaaagtt agcaccagac tttgacaaga ttgttgaaag
      601 ccttagctta ttgaaggact tttttacctc aggttctccg gaagaaacgg cgtttagagc
      661 aacagatcgt ggatctgaaa gtgacaagca ttttagaaag gtgtcagaca aagagaaaat
      721 tgaccaactt caggaagaac ttctgcacac tcagttgaag tatcagagaa tcttggaacg
      781 attagaaaag gagaacaaag aattgagaaa attagtattg cagaaagatg acaaaggcat
      841 tcatcataga aagcttaaga aatctttgat tgacatgtat tctgaagttc ttgatgttct
      901 ctctgattat gatgccagtt ataatacgca agatcatctg ccacgggttg ttgtggttgg
      961 agatcagagt gctggaaaga ctagtgtgtt ggaaatgatt gcccaagctc gaatattccc
     1021 aagaggatct ggggagatga tgacacgttc tccagttaag gtgactctga gtgaaggtcc
     1081 tcaccatgtg gccctattta aagatagttc tcgggagttt gatcttacca aagaagaaga
     1141 tcttgcagca ttaagacatg aaatagaact tcgaatgagg aaaaatgtga aagaaggctg
     1201 taccgttagc cctgagacca tatccttaaa tgtaaaaggc cctggactac agaggatggt
     1261 gcttgttgac ttaccaggtg tgattaatac tgtgacatca ggcatggctc ctgacacaaa
     1321 ggaaactatt ttcagtatca gcaaagctta catgcagaat cctaatgcca tcatactgtg
     1381 tattcaagat ggatctgtgg atgctgaacg cagtattgtt acagacttgg tcagtcaaat
     1441 ggaccctcat ggaaggagaa ccatattcgt tttgaccaaa gtagacctgg cagagaaaaa
     1501 tgtagccagt ccaagcagga ttcagcagat aattgaagga aagctcttcc caatgaaagc
     1561 tttaggttat tttgctgttg taacaggaaa agggaacagc tctgaaagca ttgaagctat
     1621 aagagaatat gaagaagagt tttttcagaa ttcaaagctc ctaaagacaa gcatgctaaa
     1681 ggcacaccaa gtgactacaa gaaatttaag ccttgcagta tcagactgct tttggaaaat
     1741 ggtacgagag tctgttgaac aacaggctga tagtttcaaa gcaacacgtt ttaaccttga
     1801 aactgaatgg aagaataact atcctcgcct gcgggaactt gaccggaatg aactatttga
     1861 aaaagctaaa aatgaaatcc ttgatgaagt tatcagtctg agccaggtta caccaaaaca
     1921 ttgggaggaa atccttcaac aatctttgtg ggaaagagta tcaactcatg tgattgaaaa
     1981 catctacctt ccagctgcgc agaccatgaa ttcaggaact tttaacacca cagtggatat
     2041 caagcttaaa cagtggactg ataaacaact tcctaataaa gcagtagagg ttgcttggga
     2101 gaccctacaa gaagaatttt cccgctttat gacagaaccg aaagggaaag agcatgatga
     2161 catatttgat aaacttaaag aggccgttaa ggaagaaagt attaaacgac acaagtggaa
     2221 tgactttgcg gaggacagct tgagggttat tcaacacaat gctttggaag accgatccgc
     2281 gtctgataaa cagcaatggg atgcagctat ttattttatg gaagaggctc tgcaggctcg
     2341 tctcaaggat actgaaaatg caattgaaaa catggtgggt ccagactgga aaaagaggtg
     2401 gttatactgg aagaatcgga cccaagaaca gtgtgttcac aatgaaacca agaatgaatt
     2461 ggagaagatg ttgaaatgta atgaggagca cccagcttat cttgcaagtg atgaaataac
     2521 cacagtccgg aagaaccttg aatcccgagg agtagaagta gatccaagct tgattaagga
     2581 tacttggcat caagtttata gaagacattt tttaaaaaca gctctaaacc attgtaacct
     2641 ttgtcgaaga ggtttttatt actaccaaag gcattttgta gattctgagt tggaatgcaa
     2701 tgatgtggtc ttgttttggc gtatacagcg catgcttgct atcaccgcaa atactttaag
     2761 gcaacaactt acaaatactg aagttaggcg attagagaaa aatgttaaag aggtattgga
     2821 agattttgct gaagatggtg agaagaagat taaattgctt actggtaaac gcgttcaact
     2881 ggcggaagac ctcaagaaag ttagagaaat tcaagaaaaa cttgatgctt tcattgaagc
     2941 tcttcatcag gagaaagggc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     3001 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3061 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3121 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3181 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3241 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3301 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3361 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3421 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3481 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3541 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     3601 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3661 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     3721 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3781 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3841 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3901 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3961 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4021 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4081 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4141 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4201 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4261 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4321 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4381 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4561 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4801 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4861 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4921 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4981 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5041 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5101 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5161 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5221 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5281 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5341 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5401 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5461 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5521 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5581 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5641 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5701 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5761 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5821 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5881 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5941 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6001 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6061 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6121 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6181 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6241 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6301 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6361 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6421 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6481 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6541 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6601 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6661 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6721 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6781 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6841 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6901 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6961 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7021 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     7141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     7201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     7261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     7321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     7381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     7441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     7501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     7561 gg
//
