LOCUS       sequence-404957-        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-4
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..37
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3000..3009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3006..3008,3009..3011,3012..3725)
                     /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(3051..3072)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3312..3331)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3659..3680
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3848..3969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3885..3904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3939..3958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3976..4431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4113..4134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4325..4344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4458..4562
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4564..4921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4564..4755
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4587..4607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4772..4907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4956..5750
                     /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             4956..5750
                     /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(5010..5029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5620..5639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5938..5957)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5982..6029
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6358..6946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6847..6866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7121..7424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7425..7562,1..66)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
       61 agagctggtt tagtgaaccg tcagatccgc tagcgctacc gatgtggcga ctacgtcggg
      121 ccgctgtggc ctgtgaggtc tgccagtctt tagtgaaaca cagctctgga ataaaaggaa
      181 gtttaccact acaaaaacta catctggttt cacgaagcat ttatcattca catcatccta
      241 ccttaaagct tcaacgaccc caattaagga catcctttca gcagttctct tctctgacaa
      301 accttccttt acgtaaactg aaattctctc caattaaata tggctaccag cctcgcagga
      361 atttttggcc agcaagatta gctacgagac tcttaaaact tcgctatctc atactaggat
      421 cggctgttgg gggtggctac acagccaaaa agacttttga tcagtggaaa gatatgatac
      481 cggaccttag tgaatataaa tggattgtgc ctgacattgt gtgggaaatt gatgagtata
      541 tcgattttga gaaaattaga aaagcccttc ctaattcaga agaccttgta aagttagcac
      601 cagactttga caagattgtt gaaagcctta gcttattgaa ggactttttt acctcaggtt
      661 ctccggaaga aacggcgttt agagcaacag atcgtggatc tgaaagtgac aagcatttta
      721 gaaaggtgtc agacaaagag aaaattgacc aacttcagga agaacttctg cacactcagt
      781 tgaagtatca gagaatcttg gaacgattag aaaaggagaa caaagaattg agaaaattag
      841 tattgcagaa agatgacaaa ggcattcatc atagaaagct taagaaatct ttgattgaca
      901 tgtattctga agttcttgat gttctctctg attatgatgc cagttataat acgcaagatc
      961 atctgccacg ggttgttgtg gttggagatc agagtgctgg aaagactagt gtgttggaaa
     1021 tgattgccca agctcgaata ttcccaagag gatctgggga gatgatgaca cgttctccag
     1081 ttaaggtgac tctgagtgaa ggtcctcacc atgtggccct atttaaagat agttctcggg
     1141 agtttgatct taccaaagaa gaagatcttg cagcattaag acatgaaata gaacttcgaa
     1201 tgaggaaaaa tgtgaaagaa ggctgtaccg ttagccctga gaccatatcc ttaaatgtaa
     1261 aaggccctgg actacagagg atggtgcttg ttgacttacc aggtgtgatt aatactgtga
     1321 catcaggcat ggctcctgac acaaaggaaa ctattttcag tatcagcaaa gcttacatgc
     1381 agaatcctaa tgccatcata ctgtgtattc aagatggatc tgtggatgct gaacgcagta
     1441 ttgttacaga cttggtcagt caaatggacc ctcatggaag gagaaccata ttcgttttga
     1501 ccaaagtaga cctggcagag aaaaatgtag ccagtccaag caggattcag cagataattg
     1561 aaggaaagct cttcccaatg aaagctttag gttattttgc tgttgtaaca ggaaaaggga
     1621 acagctctga aagcattgaa gctataagag aatatgaaga agagtttttt cagaattcaa
     1681 agctcctaaa gacaagcatg ctaaaggcac accaagtgac tacaagaaat ttaagccttg
     1741 cagtatcaga ctgcttttgg aaaatggtac gagagtctgt tgaacaacag gctgatagtt
     1801 tcaaagcaac acgttttaac cttgaaactg aatggaagaa taactatcct cgcctgcggg
     1861 aacttgaccg gaatgaacta tttgaaaaag ctaaaaatga aatccttgat gaagttatca
     1921 gtctgagcca ggttacacca aaacattggg aggaaatcct tcaacaatct ttgtgggaaa
     1981 gagtatcaac tcatgtgatt gaaaacatct accttccagc tgcgcagacc atgaattcag
     2041 gaacttttaa cgccacagtg gatatcaagc ttaaacagtg gactgataaa caacttccta
     2101 ataaagcagt agaggttgct tgggagaccc tacaagaaga attttcccgc tttatgacag
     2161 aaccgaaagg gaaagagcat gatgacatat ttgataaact taaagaggcc gttaaggaag
     2221 aaagtattaa acgacacaag tggaatgact ttgcggagga cagcttgagg gttattcaac
     2281 acaatgcttt ggaagaccga tccatatctg ataaacagca atgggatgca gctatttatt
     2341 ttatggaaga ggctctgcag gctcgtctca aggatactga aaatgcaatt gaaaacatgg
     2401 tgggtccaga ctggaaaaag aggtggttat actggaagaa tcggacccaa gaacagtgtg
     2461 ttcacaatga aaccaagaat gaattggaga agatgttgaa atgtaatgag gagcacccag
     2521 cttatcttgc aagtgatgaa ataaccacag tccggaagaa ccttgaatcc cgaggagtag
     2581 aagtagatcc aagcttgatt aaggatactt ggcatcaagt ttatagaaga cattttttaa
     2641 aaacagctct aaaccattgt aacctttgtc gaagaggttt ttattactac caaaggcatt
     2701 ttgtagattc tgagttggaa tgcaatgatg tggtcttgtt ttggcgtata cagcgcatgc
     2761 ttgctatcac cgcaaatact ttaaggcaac aacttacaaa tactgaagtt aggcgattag
     2821 agaaaaatgt taaagaggta ttggaagatt ttgctgaaga tggtgagaag aagattaaat
     2881 tgcttactgg taaacgcgtt caactggcgg aagacctcaa gaaagttaga gaaattcaag
     2941 aaaaacttga tgctttcatt gaagctcttc atcaggagaa agggctagcg ctaccggtcg
     3001 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3061 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3121 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3181 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3241 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3301 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3361 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3421 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3481 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3541 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3601 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     3661 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3721 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3781 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3841 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3961 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4021 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4081 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4141 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4201 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4261 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4321 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4381 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4441 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4501 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4561 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4621 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4741 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4861 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4921 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4981 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5041 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5101 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5161 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5221 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5281 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5341 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5401 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5461 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5521 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5581 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5641 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5701 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5761 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5821 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5881 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5941 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6001 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     6061 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     6121 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6181 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6241 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6301 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6481 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7021 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     7081 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     7141 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7201 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7261 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     7321 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     7381 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     7441 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     7501 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7561 ac
//
