LOCUS       sequence-404950-        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-8
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..56
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3019..3028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3025..3027,3028..3030,3031..3744)
                     /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(3070..3091)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3331..3350)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3678..3699
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3867..3988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3904..3923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3958..3977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3995..4450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4132..4153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4344..4363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4477..4581
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4583..4940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4583..4774
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4606..4626)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4791..4926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4975..5769
                     /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             4975..5769
                     /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(5029..5048)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5639..5658
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5957..5976)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6001..6048
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6377..6965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6866..6885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7140..7443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7444..7562,1..85)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ctttccaaaa tgtcgtaaca actccacccc attgacgcaa atgggcggta ggcgtgtacg
       61 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      121 atgtggcgac tacgtcgggc cgctgtggcc tgtgaggtct gccagtcttt agtgaaacac
      181 agctctggaa taaaaggaag tttaccacta caaaaactac atctggtttc acgaagcatt
      241 tatcattcac atcatcctac cttaaagctt caacgacccc aattaaggac atcctttcag
      301 cagttctctt ctctgacaaa ccttccttta cgtaaactga aattctctcc aattaaatat
      361 ggctaccagc ctcgcaggaa tttttggcca gcaagattag ctacgagact cttaaaactt
      421 cgctatctca tactaggatc ggctgttggg ggtggctaca cagccaaaaa gacttttgat
      481 cagtggaaag atatgatacc ggaccttagt gaatataaat ggattgtgcc tgacattgtg
      541 tgggaaattg atgagtatat cgattttgag aaaattagaa aagcccttcc taattcagaa
      601 gaccttgtaa agttagcacc agactttgac aagattgttg aaagccttag cttattgaag
      661 gactttttta cctcaggttc tccggaagaa acggcgttta gagcaacaga tcgtggatct
      721 gaaagtgaca agcattttag aaaggtgtca gacaaagaga aaattgacca acttcaggaa
      781 gaacttctgc acactcagtt gaagtatcag agaatcttgg aagcgttaga aaaggagaac
      841 aaagaattga gaaaattagt attgcagaaa gatgacaaag gcattcatca tagaaagctt
      901 aagaaatctt tgattgacat gtattctgaa gttcttgatg ttctctctga ttatgatgcc
      961 agttataata cgcaagatca tctgccacgg gttgttgtgg ttggagatca gagtgctgga
     1021 aagactagtg tgttggaaat gattgcccaa gctcgaatat tcccaagagg atctggggag
     1081 atgatgacac gttctccagt taaggtgact ctgagtgaag gtcctcacca tgtggcccta
     1141 tttaaagata gttctcggga gtttgatctt accaaagaag aagatcttgc agcattaaga
     1201 catgaaatag aacttcgaat gaggaaaaat gtgaaagaag gctgtaccgt tagccctgag
     1261 accatatcct taaatgtaaa aggccctgga ctacagagga tggtgcttgt tgacttacca
     1321 ggtgtgatta atactgtgac atcaggcatg gctcctgaca caaaggaaac tattttcagt
     1381 atcagcaaag cttacatgca gaatcctaat gccatcatac tgtgtattca agatggatct
     1441 gtggatgctg aacgcagtat tgttacagac ttggtcagtc aaatggaccc tcatggaagg
     1501 agaaccatat tcgttttgac caaagtagac ctggcagaga aaaatgtagc cagtccaagc
     1561 aggattcagc agataattga aggaaagctc ttcccaatga aagctttagg ttattttgct
     1621 gttgtaacag gaaaagggaa cagctctgaa agcattgaag ctataagaga atatgaagaa
     1681 gagttttttc agaattcaaa gctcctaaag acaagcatgc taaaggcaca ccaagtgact
     1741 acaagaaatt taagccttgc agtatcagac tgcttttgga aaatggtacg agagtctgtt
     1801 gaacaacagg ctgatagttt caaagcaaca cgttttaacc ttgaaactga atggaagaat
     1861 aactatcctc gcctgcggga acttgaccgg aatgaactat ttgaaaaagc taaaaatgaa
     1921 atccttgatg aagttatcag tctgagccag gttacaccaa aacattggga ggaaatcctt
     1981 caacaatctt tgtgggaaag agtatcaact catgtgattg aaaacatcta ccttccagct
     2041 gcgcagacca tgaattcagg aacttttaac accacagtgg atatcaagct taaacagtgg
     2101 actgataaac aacttcctaa taaagcagta gaggttgctt gggagaccct acaagaagaa
     2161 ttttcccgct ttatgacaga accgaaaggg aaagagcatg atgacatatt tgataaactt
     2221 aaagaggccg ttaaggaaga aagtattaaa cgacacaagt ggaatgactt tgcggaggac
     2281 agcttgaggg ttattcaaca caatgctttg gaagaccgat ccatatctga taaacagcaa
     2341 tgggatgcag ctatttattt tatggaagag gctctgcagg ctcgtctcaa ggatactgaa
     2401 aatgcaattg aaaacatggt gggtccagac tggaaaaaga ggtggttata ctggaagaat
     2461 cggacccaag aacagtgtgt tcacaatgaa accaagaatg aattggagaa gatgttgaaa
     2521 tgtaatgagg agcacccagc ttatcttgca agtgatgaaa taaccacagt ccggaagaac
     2581 cttgaatccc gaggagtaga agtagatcca agcttgatta aggatacttg gcatcaagtt
     2641 tatagaagac attttttaaa aacagctcta aaccattgta acctttgtcg aagaggtttt
     2701 tattactacc aaaggcattt tgtagattct gagttggaat gcaatgatgt ggtcttgttt
     2761 tggcgtatac agcgcatgct tgctatcacc gcaaatactt taaggcaaca acttacaaat
     2821 actgaagtta ggcgattaga gaaaaatgtt aaagaggtat tggaagattt tgctgaagat
     2881 ggtgagaaga agattaaatt gcttactggt aaacgcgttc aactggcgga agacctcaag
     2941 aaagttagag aaattcaaga aaaacttgat gctttcattg aagctcttca tcaggagaaa
     3001 gggctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     3061 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3121 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3181 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3241 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3301 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3361 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3421 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3481 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3541 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3601 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     3661 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3721 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3781 catttgtaga ggttttactt gctttaaaaa acatcccaca cctccccctg aacctgaaac
     3841 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3961 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4021 aaattcgcgt taaatttttg ttaaatcagc tcatttttta acaaataggc cgaaatcggc
     4081 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4141 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4201 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4261 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4321 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4381 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4441 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4561 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4621 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4681 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4741 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4801 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4861 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4921 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4981 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5041 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5101 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5161 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5221 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5281 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5341 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5401 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5461 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5521 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5581 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5641 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5701 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5761 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5821 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5881 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5941 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6001 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6061 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6121 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6181 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6961 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7021 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgaatt
     7081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgccaattg
     7201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7441 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     7501 tttccaagtc tacaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     7561 ga
//
