LOCUS       sequence-404949-        7562 bp    DNA     circular SYN 15-APR-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 Apr 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-7
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2972..2981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2978..2980,2981..2983,2984..3697)
                     /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(3023..3044)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3284..3303)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3631..3652
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3820..3941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3857..3876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3911..3930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3948..4403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4085..4106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4297..4316
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4430..4534
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4536..4893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4536..4727
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4559..4579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4744..4879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4928..5722
                     /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             4928..5722
                     /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(4982..5001)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5592..5611
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5910..5929)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5954..6001
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6330..6918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6819..6838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7093..7396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7397..7562,1..38)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7551..7562,1..9)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
       61 gctagcgcta ccgatgtggc gactacgtcg ggccgctgtg gcctgtgagg tctgccagtc
      121 tttagtgaaa cacagctctg gaataaaagg aagtttacca ctacaaaaac tacatctggt
      181 ttcacgaagc atttatcatt cacatcatcc taccttaaag cttcaacgac cccaattaag
      241 gacatccttt cagcagttct cttctctgac aaaccttcct ttacgtaaac tgaaattctc
      301 tccaattaaa tatggctacc agcctcgcag gaatttttgg ccagcaagat tagctacgag
      361 actcttaaaa cttcgctatc tcatactagg atcggctgtt gggggtggct acacagccaa
      421 aaagactttt gatcagtgga aagatatgat accggacctt agtgaatata aatggattgt
      481 gcctgacatt gtgtgggaaa ttgatgagta tatcgatttt gagaaaatta gaaaagccct
      541 tcctaattca gaagaccttg taaagttagc accagacttt gacaagattg ttgaaagcct
      601 tagcttattg aaggactttt ttacctcagg ttctccggaa gaaacggcgt ttagagcaac
      661 agatcgtgga tctgaaagtg acaagcattt tagaaaggtg tcagacaaag agaaaattga
      721 ccaacttcag gaagaacttc tgcacactca gttgaagtat cagagaatct tggaacgatt
      781 agaaaaggag aacaaagaat tgagaaaatt agtattgcag aaagatgaca aaggcattca
      841 tcatagaaag cttaagaaat ctttgattga catgtattct gaagttcttg atgttctctc
      901 tgattatgat gccagttata atacgcaaga tcatctgcca cgggttgttg tggttggaga
      961 tcagagtgct ggaaagacta gtgtgttgga aatgattgcc caagctcgaa tattcccaag
     1021 aggatctggg gagatgatga cacgttctcc agttaaggtg actctgagtg aaggtcctca
     1081 ccatgtggcc ctatttaaag atagttctcg ggagtttgat cttaccaaag aagaagatct
     1141 tgcagcatta agacatgaaa tagaacttcg aatgaggaaa aatgtgaaag aaggctgtac
     1201 cgttagccct gagaccatat ccttaaatgt aaaaggccct ggactacaga ggatggtgct
     1261 tgttgactta ccaggtgtga ttaatactgt gacatcaggc atggctcctg acacaaagga
     1321 aactattttc agtatcagca aagcttacat gcagaatcct aatgccatca tactgtgtat
     1381 tcaagatgga tctgtggatg ctgaacgcag tattgttaca gacttggtca gtcaaatgga
     1441 ccctcatgga aggagaacca tattcgtttt gaccaaagta gacctggcag agaaaaatgt
     1501 agccagtcca agcaggattc agcagataat tgaaggaaag ctcttcccaa tgaaagcttt
     1561 aggttatttt gctgttgtaa caggaaaagg gaacagctct gaaagcattg aagctataag
     1621 agaatatgaa gaagagtttt ttcagaattc aaagctccta aagacaagca tgctaaaggc
     1681 acaccaagtg actacaagaa atttaagcct tgcagtatca gactgctttt ggaaaatggt
     1741 acgagagtct gttgaacaac aggctgatag tttcaaagca acacgtttta accttgaaac
     1801 tgaatggaag aataactatc ctcgcctgcg ggaacttgac cggaatgaac tatttgaaaa
     1861 agctaaaaat gaaatccttg atgaagttat cagtctgagc caggttacac caaaacattg
     1921 ggaggaaatc cttcaacaat ctttgtggga aagagtatca actcatgtga ttgaaaacat
     1981 ctaccttcca gctgcgcaga ccatgaattc aggaactttt aacaccacag tggatatcaa
     2041 gcttaaacag tggactgata aacaacttcc taataaagca gtagaggttg cttgggagac
     2101 cctacaagaa gaattttccc gctttatgac agaaccgaaa gggaaagagc atgatgacat
     2161 atttgataaa cttaaagagg ccgttaagga agaaagtatt aaacgacaca agtggaatga
     2221 ctttgcggag gacagcttga gggttattca acacaatgct ttggaagacc gatccatatc
     2281 tgataaacag caatgggatg cagctattta ttttatggaa gaggctctgc aggctcgtct
     2341 caaggatact gaaaatgcaa ttgaaaacat ggtgggtcca gactggaaaa agaggtggtt
     2401 atactggaag aatcggaccc aagaacagtg tgttcacaat gaaaccaaga atgaattgga
     2461 gaagatgttg aaatgtaatg aggagcaccc agcttatctt gcaagtgatg aaataaccac
     2521 agtccgggcg gcccttgaat cccgaggagt agaagtagat ccaagcttga ttaaggatac
     2581 ttggcatcaa gtttatagaa gacatttttt aaaaacagct ctaaaccatt gtaacctttg
     2641 tcgaagaggt ttttattact accaaaggca ttttgtagat tctgagttgg aatgcaatga
     2701 tgtggtcttg ttttggcgta tacagcgcat gcttgctatc accgcaaata ctttaaggca
     2761 acaacttaca aatactgaag ttaggcgatt agagaaaaat gttaaagagg tattggaaga
     2821 ttttgctgaa gatggtgaga agaagattaa attgcttact ggtaaacgcg ttcaactggc
     2881 ggaagacctc aagaaagtta gagaaattca agaaaaactt gatgctttca ttgaagctct
     2941 tcatcaggag aaagggctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     3001 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3061 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3121 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3181 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3241 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3301 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3361 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3421 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3481 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3541 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     3601 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3661 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     3721 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3781 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3901 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3961 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4021 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4081 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4141 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4201 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4261 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4321 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4381 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4441 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4501 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4561 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4621 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4741 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4801 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4861 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4921 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4981 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5041 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5101 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5161 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5221 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5281 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5341 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5401 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5461 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5521 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5581 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5641 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5701 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5761 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5821 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5881 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5941 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6001 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6061 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6121 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     6181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6961 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7021 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     7081 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7141 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7201 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7261 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7321 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7381 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7441 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7501 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     7561 cg
//
