LOCUS       sequence-404940-        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-2
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2974..2983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2980..2982,2983..2985,2986..3699)
                     /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(3025..3046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3286..3305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3633..3654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3822..3943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3859..3878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3913..3932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3950..4405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4087..4108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4299..4318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4432..4536
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4538..4895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4538..4729
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4561..4581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4746..4881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4930..5724
                     /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             4930..5724
                     /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(4984..5003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5594..5613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5912..5931)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5956..6003
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6332..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7095..7398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7399..7562,1..40)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7553..7562,1..11)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
       61 ccgctagcgc taccgatgtg gcgactacgt cgggccgctg tggcctgtga ggtctgccag
      121 tctttagtga aacacagctc tggaataaaa ggaagtttac cactacaaaa actacatctg
      181 gtttcacgaa gcatttatca ttcacatcat cctaccttaa agcttcaacg accccaatta
      241 aggacatcct ttcagcagtt ctcttctctg acaaaccttc ctttacgtaa actgaaattc
      301 tctccaatta aatatggcta ccagcctcgc aggaattttt ggccagcaag attagctacg
      361 agactcttaa aacttcgcta tctcatacta ggatcggctg ttgggggtgg ctacacagcc
      421 aaaaagactt ttgatcagtg gaaagatatg ataccggacc ttagtgaata taaatggatt
      481 gtgcctgaca ttgtgtggga aattgatgag tatatcgatt ttgagaaaat tagaaaagcc
      541 cttcctaatt cagaagacct tgtaaagtta gcaccagact ttgacaagat tgttgaaagc
      601 cttagcttat tgaaggactt ttttacctca ggttctccgg aagaaacggc gtttagagca
      661 acagatcgtg gatctgaaag tgacaagcat tttagaaagg tgtcagacaa agagaaaatt
      721 gaccaacttc aggaagaact tctgcacact cagttgaagt atcagagaat cttggaacga
      781 ttagaaaagg agaacaaaga attgagaaaa ttagtattgc agaaagatga caaaggcatt
      841 catcatagaa agcttaagaa atctttgatt gacatgtatt ctgaagttct tgatgttctc
      901 tctgattatg atgccagtta taatacgcaa gatcatctgc cacgggttgt tgtggttgga
      961 gatcagagtg ctggaaagac tagtgtgttg gaaatgattg cccaagctcg aatattccca
     1021 agaggatctg gggagatgat gacacgttct ccagttaagg tgactctgag tgaaggtcct
     1081 caccatgtgg ccctatttaa agatagttct cgggagtttg atcttaccaa agaagaagat
     1141 cttgcagcat taagacatga aatagaactt cgaatgagga aaaatgtgaa agaaggctgt
     1201 accgttagcc ctgagaccat atccttaaat gtaaaaggcc ctggactaca gaggatggtg
     1261 cttgttgact taccaggtgt gattaatact gtgacatcag gcatggctcc tgacacaaag
     1321 gaaactattt tcagtatcag caaagcttac atgcagaatc ctaatgccat catactgtgt
     1381 attcaagatg gatctgtgga tgctgaacgc agtattgtta cagacttggt cagtcaaatg
     1441 gaccctcatg gaaggagaac catattcgtt ttgaccaaag tagacctggc agagaaaaat
     1501 gtagccagtc caagcaggat tcagcagata attgaaggaa agctcttccc aatgaaagct
     1561 ttaggttatt ttgctgttgt aacaggaaaa gggaacagct ctgaaagcat tgaagctata
     1621 agagaatatg aagaagagtt ttttcagaat tcaaagctcc taaagacaag catgctaaag
     1681 gcacaccaag tgactacaag aaatttaagc cttgcagtat cagactgctt ttggaaaatg
     1741 gtacgagagt ctgttgaaca acaggctgat agtttcaaag caacacgttt taaccttgaa
     1801 actgaatgga agaataacta tcctcgcctg cgggaacttg accggaatga actatttgaa
     1861 aaagctaaaa atgaaatcct tgatgaagtt atcagtctga gccaggttac accagcagct
     1921 tgggaggaaa tccttcaaca atctttgtgg gaaagagtat caactcatgt gattgaaaac
     1981 atctaccttc cagctgcgca gaccatgaat tcaggaactt ttaacaccac agtggatatc
     2041 aagcttaaac agtggactga taaacaactt cctaataaag cagtagaggt tgcttgggag
     2101 accctacaag aagaattttc ccgctttatg acagaaccga aagggaaaga gcatgatgac
     2161 atatttgata aacttaaaga ggccgttaag gaagaaagta ttaaacgaca caagtggaat
     2221 gactttgcgg aggacagctt gagggttatt caacacaatg ctttggaaga ccgatccata
     2281 tctgataaac agcaatggga tgcagctatt tattttatgg aagaggctct gcaggctcgt
     2341 ctcaaggata ctgaaaatgc aattgaaaac atggtgggtc cagactggaa aaagaggtgg
     2401 ttatactgga agaatcggac ccaagaacag tgtgttcaca atgaaaccaa gaatgaattg
     2461 gagaagatgt tgaaatgtaa tgaggagcac ccagcttatc ttgcaagtga tgaaataacc
     2521 acagtccgga agaaccttga atcccgagga gtagaagtag atccaagctt gattaaggcg
     2581 acttggcatc aagtttatag aagacatttt ttaaaaacag ctctaaacca ttgtaacctt
     2641 tgtcgaagag gtttttatta ctaccaaagg cattttgtag attctgagtt ggaatgcaat
     2701 gatgtggtct tgttttggcg tatacagcgc atgcttgcta tcaccgcaaa tactttaagg
     2761 caacaactta caaatactga agttaggcga ttagagaaaa atgttaaaga ggtattggaa
     2821 gattttgctg aagatggtga gaagaagatt aaattgctta ctggtaaacg cgttcaactg
     2881 gcggaagacc tcaagaaagt tagagaaatt caagaaaaac ttgatgcttt cattgaagct
     2941 cttcatcagg agaaagggct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     3001 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3061 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3121 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3181 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3241 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3301 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3361 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3421 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3481 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3541 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     3601 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3661 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     3721 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3781 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3841 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3901 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3961 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4021 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4081 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4141 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4201 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4261 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4321 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4381 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4501 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4621 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4801 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4861 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4921 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4981 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5041 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5101 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5161 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5221 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5281 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5341 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5401 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5461 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5521 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5581 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5641 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5701 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5761 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5821 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5881 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5941 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     6001 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6061 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6121 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6181 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6241 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6301 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6361 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6421 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7021 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     7081 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     7141 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     7201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     7261 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     7321 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     7381 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     7441 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     7501 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     7561 gg
//
