LOCUS       sequence-404951-        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-9
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..58
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3021..3030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3027..3029,3030..3032,3033..3746)
                     /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(3072..3093)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3333..3352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3680..3701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3869..3990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3906..3925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3960..3979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3997..4452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4134..4155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4346..4365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4479..4583
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4585..4942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4585..4776
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4608..4628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4793..4928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4977..5771
                     /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             4977..5771
                     /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(5031..5050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5641..5660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5959..5978)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6003..6050
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6379..6967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6868..6887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7142..7445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7446..7562,1..87)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
       61 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      121 cgatgtggcg actacgtcgg gccgctgtgg cctgtgaggt ctgccagtct ttagtgaaac
      181 acagctctgg aataaaagga agtttaccac tacaaaaact acatctggtt tcacgaagca
      241 tttatcattc acatcatcct accttaaagc ttcaacgacc ccaattaagg acatcctttc
      301 agcagttctc ttctctgaca aaccttcctt tacgtaaact gaaattctct ccaattaaat
      361 atggctacca gcctcgcagg aatttttggc cagcaagatt agctacgaga ctcttaaaac
      421 ttcgctatct catactagga tcggctgttg ggggtggcta cacagccaaa aagacttttg
      481 atcagtggaa agatatgata ccggacctta gtgaatataa atggattgtg cctgacattg
      541 tgtgggaaat tgatgagtat atcgattttg agaaaattag aaaagccctt cctaattcag
      601 aagaccttgt aaagttagca ccagactttg acaagattgt tgaaagcctt agcttattga
      661 aggacttttt tacctcaggt tctccggaag aaacggcgtt tagagcaaca gatcgtggat
      721 ctgaaagtga caagcatttt agaaaggtgt cagacaaagc gaaaattgac gcacttcagg
      781 aagaacttct gcacactcag ttgaagtatc agagaatctt ggaacgatta gaaaaggaga
      841 acaaagaatt gagaaaatta gtattgcaga aagatgacaa aggcattcat catagaaagc
      901 ttaagaaatc tttgattgac atgtattctg aagttcttga tgttctctct gattatgatg
      961 ccagttataa tacgcaagat catctgccac gggttgttgt ggttggagat cagagtgctg
     1021 gaaagactag tgtgttggaa atgattgccc aagctcgaat attcccaaga ggatctgggg
     1081 agatgatgac acgttctcca gttaaggtga ctctgagtga aggtcctcac catgtggccc
     1141 tatttaaaga tagttctcgg gagtttgatc ttaccaaaga agaagatctt gcagcattaa
     1201 gacatgaaat agaacttcga atgaggaaaa atgtgaaaga aggctgtacc gttagccctg
     1261 agaccatatc cttaaatgta aaaggccctg gactacagag gatggtgctt gttgacttac
     1321 caggtgtgat taatactgtg acatcaggca tggctcctga cacaaaggaa actattttca
     1381 gtatcagcaa agcttacatg cagaatccta atgccatcat actgtgtatt caagatggat
     1441 ctgtggatgc tgaacgcagt attgttacag acttggtcag tcaaatggac cctcatggaa
     1501 ggagaaccat attcgttttg accaaagtag acctggcaga gaaaaatgta gccagtccaa
     1561 gcaggattca gcagataatt gaaggaaagc tcttcccaat gaaagcttta ggttattttg
     1621 ctgttgtaac aggaaaaggg aacagctctg aaagcattga agctataaga gaatatgaag
     1681 aagagttttt tcagaattca aagctcctaa agacaagcat gctaaaggca caccaagtga
     1741 ctacaagaaa tttaagcctt gcagtatcag actgcttttg gaaaatggta gcagagtctg
     1801 ttgaacaaca ggctgcgagt ttcaaagcaa cacgttttaa ccttgaaact gaatggaaga
     1861 ataactatcc tcgcctgcgg gaacttgacc ggaatgaact atttgaaaaa gctaaaaatg
     1921 aaatccttga tgaagttatc agtctgagcc aggttacacc aaaacattgg gaggaaatcc
     1981 ttcaacaatc tttgtgggaa agagtatcaa ctcatgtgat tgaaaacatc taccttccag
     2041 ctgcgcagac catgaattca ggaactttta acaccacagt ggatatcaag cttaaacagt
     2101 ggactgataa acaacttcct aataaagcag tagaggttgc ttgggagacc ctacaagaag
     2161 aattttcccg ctttatgaca gaaccgaaag ggaaagagca tgatgacata tttgataaac
     2221 ttaaagaggc cgttaaggaa gaaagtatta aacgacacaa gtggaatgac tttgcggagg
     2281 acagcttgag ggttattcaa cacaatgctt tggaagaccg atccatatct gataaacagc
     2341 aatgggatgc agctatttat tttatggaag aggctctgca ggctcgtctc aaggatactg
     2401 aaaatgcaat tgaaaacatg gtgggtccag actggaaaaa gaggtggtta tactggaaga
     2461 atcggaccca agaacagtgt gttcacaatg aaaccaagaa tgaattggag aagatgttga
     2521 aatgtaatga ggagcaccca gcttatcttg caagtgatga aataaccaca gtccggaaga
     2581 accttgaatc ccgaggagta gaagtagatc caagcttgat taaggatact tggcatcaag
     2641 tttatagaag acatttttta aaaacagctc taaaccattg taacctttgt cgaagaggtt
     2701 tttattacta ccaaaggcat tttgtagatt ctgagttgga atgcaatgat gtggtcttgt
     2761 tttggcgtat acagcgcatg cttgctatca ccgcaaatac tttaaggcaa caacttacaa
     2821 atactgaagt taggcgatta gagaaaaatg ttaaagaggt attggaagat tttgctgaag
     2881 atggtgagaa gaagattaaa ttgcttactg gtaaacgcgt tcaactggcg gaagacctca
     2941 agaaagttag agaaattcaa gaaaaacttg atgctttcat tgaagctctt catcaggaga
     3001 aagggctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3061 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3121 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3181 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3241 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3301 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3361 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3421 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3481 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3541 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3601 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     3661 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3721 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     3781 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3841 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3901 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3961 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4021 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4081 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4201 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4261 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4321 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4381 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4441 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4501 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4561 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4621 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4681 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4741 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4861 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4921 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4981 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5041 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5101 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5161 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5221 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5281 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5341 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5401 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5461 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5521 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5581 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5641 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5701 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5761 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5821 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5881 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5941 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6001 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6061 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6121 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6181 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6241 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7081 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     7141 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     7201 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     7261 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     7321 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     7381 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     7441 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     7501 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     7561 gg
//
