LOCUS       sequence-404954-        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-1
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     82..102
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3065..3074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3071..3073,3074..3076,3077..3790)
                     /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(3116..3137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3377..3396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3724..3745
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3913..4034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4004..4023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4041..4496)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4178..4199)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4390..4409
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4523..4627
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4629..4986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4629..4820
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4652..4672)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4837..4972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5021..5815
                     /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             5021..5815
                     /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(5075..5094)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5685..5704
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6003..6022)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6047..6094
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6423..7011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6912..6931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7186..7489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7490..7562,1..131)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
       61 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      121 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccgatgt ggcgactacg
      181 tcgggccgct gtggcctgtg aggtctgcca gtctttagtg aaacacagct ctggaataaa
      241 aggaagttta ccactacaaa aactacatct ggtttcacga agcatttatc attcacatca
      301 tcctacctta aagcttcaac gaccccaatt aaggacatcc tttcagcagt tctcttctct
      361 gacaaacctt cctttacgta aactgaaatt ctctccaatt aaatatggct accagcctcg
      421 caggaatttt tggccagcaa gattagctac gagactctta aaacttcgct atctcatact
      481 aggatcggct gttgggggtg gctacacagc caaaaagact tttgatcagt ggaaagatat
      541 gataccggac cttagtgaat ataaatggat tgtgcctgac attgtgtggg aaattgatga
      601 gtatatcgat tttgagaaaa ttagaaaagc ccttcctaat tcagaagacc ttgtaaagtt
      661 agcaccagac tttgacaaga ttgttgaaag ccttagctta ttgaaggact tttttacctc
      721 aggttctccg gaagaaacgg cgtttagagc aacagatcgt ggatctgaaa gtgacaagca
      781 ttttagaaag gtgtcagaca aagagaaaat tgaccaactt caggaagaac ttctgcacac
      841 tcagttgaag tatcagagaa tcttggaacg attagaaaag gagaacaaag aagccagaaa
      901 attagtattg cagaaagatg acaaaggcat tcatcataga aagcttaaga aatctttgat
      961 tgacatgtat tctgaagttc ttgatgttct ctctgattat gatgccagtt ataatacgca
     1021 agatcatctg ccacgggttg ttgtggttgg agatcagagt gctggaaaga ctagtgtgtt
     1081 ggaaatgatt gcccaagctc gaatattccc aagaggatct ggggagatga tgacacgttc
     1141 tccagttaag gtgactctga gtgaaggtcc tcaccatgtg gccctattta aagatagttc
     1201 tcgggagttt gatcttacca aagaagaaga tcttgcagca ttaagacatg aaatagaact
     1261 tcgaatgagg aaaaatgtga aagaaggctg taccgttagc cctgagacca tatccttaaa
     1321 tgtaaaaggc cctggactac agaggatggt gcttgttgac ttaccaggtg tgattaatac
     1381 tgtgacatca ggcatggctc ctgacacaaa ggaaactatt ttcagtatca gcaaagctta
     1441 catgcagaat cctaatgcca tcatactgtg tattcaagat ggatctgtgg atgctgaacg
     1501 cagtattgtt acagacttgg tcagtcaaat ggaccctcat ggaaggagaa ccatattcgt
     1561 tttgaccaaa gtagacctgg cagagaaaaa tgtagccagt ccaagcagga ttcagcagat
     1621 aattgaagga aagctcttcc caatgaaagc tttaggttat tttgctgttg taacaggaaa
     1681 agggaacagc tctgaaagca ttgaagctat aagagaatat gaagaagagt tttttcagaa
     1741 ttcaaagctc ctaaagacaa gcatgctaaa ggcacaccaa gtgactacaa gaaatttaag
     1801 ccttgcagta tcagactgct tttggaaaat ggtacgagag tctgttgaac aacaggctga
     1861 tagtttcaaa gcaacacgtt ttaaccttga aactgaatgg aagaataact atcctcgcct
     1921 gcgggaactt gaccggaatg aactatttga aaaagctaaa aatgaaatcc ttgatgaagt
     1981 tatcagtctg agccaggtta caccaaaaca ttgggaggaa atccttcaac aatctttgtg
     2041 ggaaagagta tcaactcatg tgattgaaaa catctacctt ccagctgcgc agaccatgaa
     2101 ttcaggaact tttaacacca cagtggatat caagcttaaa cagtggactg ataaacaact
     2161 tcctaataaa gcagtagagg ttgcttggga gaccctacaa gaagaatttt cccgctttat
     2221 gacagaaccg aaagggaaag agcatgatga catatttgat aaacttaaag aggccgttaa
     2281 ggaagaaagt attaaacgac acaagtggaa tgactttgcg gaggacagct tgagggttat
     2341 tcaacacaat gctttggaag accgatccat atctgataaa cagcaatggg atgcagctat
     2401 ttattttatg gaagaggctc tgcaggctcg tctcaaggat actgaaaatg caattgaaaa
     2461 catggtgggt ccagactgga aaaagaggtg gttatactgg aagaatcgga cccaagaaca
     2521 gtgtgttcac aatgaaacca agaatgaatt ggagaagatg ttgaaatgta atgaggagca
     2581 cccagcttat cttgcaagtg atgaaataac cacagtccgg aagaaccttg aatcccgagg
     2641 agtagaagta gatccaagct tgattaagga tacttggcat caagtttata gaagacattt
     2701 tttaaaaaca gctctaaacc attgtaacct ttgtcgaaga ggtttttatt actaccaaag
     2761 gcattttgta gattctgagt tggaatgcaa tgatgtggtc ttgttttggc gtatacagcg
     2821 catgcttgct atcaccgcaa atactttaag gcaacaactt acaaatactg aagttaggcg
     2881 attagagaaa aatgttaaag aggtattgga agattttgct gaagatggtg agaagaagat
     2941 taaattgctt actggtaaac gcgttcaact ggcggaagac ctcaagaaag ttagagaaat
     3001 tcaagaaaaa cttgatgctt tcattgaagc tcttcatcag gagaaagggc tagcgctacc
     3061 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3181 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3241 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3361 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3421 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3481 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3541 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3661 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     3721 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3781 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     3841 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3901 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3961 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4021 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4081 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4141 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4201 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4261 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4321 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4381 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4441 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     4501 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4561 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4621 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4681 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4741 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4801 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4861 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4921 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4981 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5041 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5101 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5161 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     5221 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5281 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5341 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5401 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5461 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5521 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     5581 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5641 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5701 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5761 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5821 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5881 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5941 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     6001 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     6061 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     6121 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     6181 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     6241 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     6301 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6541 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7081 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     7141 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7201 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7261 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7321 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7381 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     7441 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     7501 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     7561 cc
//
