LOCUS       sequence-404935-        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-1 (a)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2969..2978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2975..2977,2978..2980,2981..3694)
                     /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(3020..3041)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3281..3300)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3628..3649
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3817..3938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3854..3873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3908..3927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3945..4400)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4082..4103)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4294..4313
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4427..4531
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4533..4890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4533..4724
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4556..4576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4741..4876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4925..5719
                     /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             4925..5719
                     /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(4979..4998)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5589..5608
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5907..5926)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5951..5998
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6327..6915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6816..6835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7090..7393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7394..7562,1..35)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(7548..7562,1..6)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
       61 agcgctaccg atgtggcgac tacgtcgggc cgctgtggcc tgtgaggtct gccagtcttt
      121 agtgaaacac agctctggaa taaaaggaag tttaccacta caaaaactac atctggtttc
      181 acgaagcatt tatcattcac atcatcctac cttaaagctt caacgacccc aattaaggac
      241 atcctttcag cagttctctt ctctgacaaa ccttccttta cgtaaactga aattctctcc
      301 aattaaatat ggctaccagc ctcgcaggaa tttttggcca gcaagattag ctacgagact
      361 cttaaaactt cgctatctca tactaggatc ggctgttggg ggtggctaca cagccaaaaa
      421 gacttttgat cagtggaaag atatgatacc ggaccttagt gaatataaat ggattgtgcc
      481 tgacattgtg tgggaaattg atgagtatat cgattttgag aaaattagaa aagcccttcc
      541 taattcagaa gaccttgtaa agttagcacc agactttgac aagattgttg aaagccttag
      601 cttattgaag gactttttta cctcaggttc tccggaagaa acggcgttta gagcaacaga
      661 tcgtggatct gaaagtgaca agcattttag aaaggtgtca gacaaagaga aaattgacca
      721 acttcaggaa gaacttctgc acactcagtt gaagtatcag agaatcttgg aacgattaga
      781 aaaggagaac aaagaattga gaaaattagt attgcagaaa gatgacaaag gcattcatca
      841 tagaaagctt aagaaatctt tgattgacat gtattctgaa gttcttgatg ttctctctga
      901 ttatgatgcc agttataata cgcaagatca tctgccacgg gttgttgtgg ttggagatca
      961 gagtgctgga aagactagtg tgttggaaat gattgcccaa gctcgaatat tcccaagagg
     1021 atctggggag atgatgacac gttctccagt taaggtgact ctgagtgaag gtcctcacca
     1081 tgtggcccta tttaaagata gttctcggga gtttgatctt accaaagaag aagatcttgc
     1141 agcattaaga catgaaatag aacttcgaat gaggaaaaat gtgaaagaag gctgtaccgt
     1201 tagccctgag accatatcct taaatgtaaa aggccctgga ctacagagga tggtgcttgt
     1261 tgacttacca ggtgtgatta atactgtgac atcaggcatg gctcctgaca caaaggaaac
     1321 tattttcagt atcagcaaag cttacatgca gaatcctaat gccatcatac tgtgtattca
     1381 agatggatct gtggatgctg aacgcagtat tgttacagac ttggtcagtc aaatggaccc
     1441 tcatggaagg agaaccatat tcgttttgac caaagtagac ctggcagaga aaaatgtagc
     1501 cagtccaagc aggattcagc agataattga aggaaagctc ttcccaatga aagctttagg
     1561 ttattttgct gttgtaacag gaaaagggaa cagctctgaa agcattgaag ctataagaga
     1621 atatgaagaa gagttttttc agaattcaaa gctcctaaag acaagcatgc taaaggcaca
     1681 ccaagtgact acaagaaatt taagccttgc agtatcagac tgcttttgga aaatggtacg
     1741 agagtctgtt gaacaacagg ctgatagttt caaagcaaca cgttttaacc ttgaaactga
     1801 atggaagaat aactatcctc gcctgcggga acttgaccgg aatgaactat ttgaaaaagc
     1861 taaaaatgaa atccttgatg aagttatcag tctgagccag gttacaccaa aacattggga
     1921 ggaaatcctt caacaatctt tgtgggaagc agtatcaact catgtgattg aaaacatcta
     1981 ccttccagct gcgcagacca tgaattcagg aacttttaac accacagtgg atatcaagct
     2041 taaacagtgg actgatgcac aacttcctaa taaagcagta gaggttgctt gggagaccct
     2101 acaagaagaa ttttccgcgt ttatgacaga accgaaaggg aaagagcatg atgacatatt
     2161 tgataaactt aaagaggccg ttaaggaaga aagtattaaa cgacacaagt ggaatgactt
     2221 tgcggaggac agcttgaggg ttattcaaca caatgctttg gaagaccgat ccatatctga
     2281 taaacagcaa tgggatgcag ctatttattt tatggaagag gctctgcagg ctcgtctcaa
     2341 ggatactgaa aatgcaattg aaaacatggt gggtccagac tggaaaaaga ggtggttata
     2401 ctggaagaat cggacccaag aacagtgtgt tcacaatgaa accaagaatg aattggagaa
     2461 gatgttgaaa tgtaatgagg agcacccagc ttatcttgca agtgatgaaa taaccacagt
     2521 ccggaagaac cttgaatccc gaggagtaga agtagatcca agcttgatta aggatacttg
     2581 gcatcaagtt tatagaagac attttttaaa aacagctcta aaccattgta acctttgtcg
     2641 aagaggtttt tattactacc aaaggcattt tgtagattct gagttggaat gcaatgatgt
     2701 ggtcttgttt tggcgtatac agcgcatgct tgctatcacc gcaaatactt taaggcaaca
     2761 acttacaaat actgaagtta ggcgattaga gaaaaatgtt aaagaggtat tggaagattt
     2821 tgctgaagat ggtgagaaga agattaaatt gcttactggt aaacgcgttc aactggcgga
     2881 agacctcaag aaagttagag aaattcaaga aaaacttgat gctttcattg aagctcttca
     2941 tcaggagaaa gggctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt
     3001 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3061 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3121 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3181 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3241 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3301 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3361 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3421 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3481 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3541 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     3601 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3661 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     3721 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3781 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3841 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3901 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3961 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4021 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4081 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4141 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4201 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4261 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4321 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4381 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4441 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4501 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     4561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4741 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4801 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4861 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4921 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4981 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5041 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5101 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5161 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5221 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5281 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5341 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5401 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5461 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5521 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5581 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5641 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5701 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5761 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5821 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5881 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5941 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6001 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6061 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6121 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     6181 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6241 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6301 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6361 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6421 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6481 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6541 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6601 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6661 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6721 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6781 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6841 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6901 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6961 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7021 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     7081 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7141 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7201 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7261 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7321 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     7381 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     7441 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     7501 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     7561 ta
//
