LOCUS       sequence-404936-        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 (b)
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2931..2940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2937..2939,2940..2942,2943..3656)
                     /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(2982..3003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3243..3262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3590..3611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3779..3900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3816..3835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3870..3889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3907..4362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4044..4065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4256..4275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4389..4493
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4495..4852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4495..4686
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4518..4538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4703..4838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4887..5681
                     /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             4887..5681
                     /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(4941..4960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5551..5570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5869..5888)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5913..5960
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6289..6877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6778..6797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7052..7355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7356..7559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7510..7530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ttagtgaacc gtcagatccg ctagcgctac cgatgtggcg actacgtcgg gccgctgtgg
       61 cctgtgaggt ctgccagtct ttagtgaaac acagctctgg aataaaagga agtttaccac
      121 tacaaaaact acatctggtt tcacgaagca tttatcattc acatcatcct accttaaagc
      181 ttcaacgacc ccaattaagg acatcctttc agcagttctc ttctctgaca aaccttcctt
      241 tacgtaaact gaaattctct ccaattaaat atggctacca gcctcgcagg aatttttggc
      301 cagcaagatt agctacgaga ctcttaaaac ttcgctatct catactagga tcggctgttg
      361 ggggtggcta cacagccaaa aagacttttg atcagtggaa agatatgata ccggacctta
      421 gtgaatataa atggattgtg cctgacattg tgtgggaaat tgatgagtat atcgattttg
      481 agaaaattag aaaagccctt cctaattcag aagaccttgt aaagttagca ccagactttg
      541 acaagattgt tgaaagcctt agcttattga aggacttttt tacctcaggt tctccggaag
      601 aaacggcgtt tagagcaaca gatcgtggat ctgaaagtga caagcatttt agaaaggtgt
      661 cagacaaaga gaaaattgac caacttcagg aagaacttct gcacactcag ttgaagtatc
      721 agagaatctt ggaacgatta gaaaaggaga acaaagaatt gagaaaatta gtattgcaga
      781 aagatgacaa aggcattcat catagaaagc ttaagaaatc tttgattgac atgtattctg
      841 aagttcttga tgttctctct gattatgatg ccagttataa tacgcaagat catctgccac
      901 gggttgttgt ggttggagat cagagtgctg gaaagactag tgtgttggaa atgattgccc
      961 aagctcgaat attcccaaga ggatctgggg agatgatgac acgttctcca gttaaggtga
     1021 ctctgagtga aggtcctcac catgtggccc tatttaaaga tagttctcgg gagtttgatc
     1081 ttaccaaaga agaagatctt gcagcattaa gacatgaaat agaacttcga atgaggaaaa
     1141 atgtgaaaga aggctgtacc gttagccctg agaccatatc cttaaatgta aaaggccctg
     1201 gactacagag gatggtgctt gttgacttac caggtgtgat taatactgtg acatcaggca
     1261 tggctcctga cacaaaggaa actattttca gtatcagcaa agcttacatg cagaatccta
     1321 atgccatcat actgtgtatt caagatggat ctgtggatgc tgaacgcagt attgttacag
     1381 acttggtcag tcaaatggac cctcatggaa ggagaaccat attcgttttg accaaagtag
     1441 acctggcaga gaaaaatgta gccagtccaa gcaggattca gcagataatt gaaggaaagc
     1501 tcttcccaat gaaagcttta ggttattttg ctgttgtaac aggaaaaggg aacagctctg
     1561 aaagcattga agctataaga gaatatgaag aagagttttt tcagaattca aagctcctaa
     1621 agacaagcat gctaaaggca caccaagtga ctacaagaaa tttaagcctt gcagtatcag
     1681 actgcttttg gaaaatggta cgagagtctg ttgaacaaca ggctgatagt ttcaaagcaa
     1741 cacgttttaa ccttgaaact gaatggaaga ataactatcc tcgcctgcgg gaacttgacc
     1801 ggaatgaact atttgaaaaa gctaaaaatg aaatccttga tgaagttatc agtctgagcc
     1861 aggttacacc aaaacattgg gaggaaatcc ttcaacaatc tttgtgggca agagtatcag
     1921 ctcatgtgat tgaaaacatc taccttccag ctgcgcagac catgaattca ggaactttta
     1981 acaccacagt ggatatcaag cttaaacagt ggactgataa acaacttcct aatgcagcag
     2041 tagcggttgc ttgggagacc ctacaagaag aattttcccg ctttatgaca gaaccgaaag
     2101 ggaaagagca tgatgacata tttgataaac ttaaagaggc cgttaaggaa gaaagtatta
     2161 aacgacacaa gtggaatgac tttgcggagg acagcttgag ggttattcaa cacaatgctt
     2221 tggaagaccg atccatatct gataaacagc aatgggatgc agctatttat tttatggaag
     2281 aggctctgca ggctcgtctc aaggatactg aaaatgcaat tgaaaacatg gtgggtccag
     2341 actggaaaaa gaggtggtta tactggaaga atcggaccca agaacagtgt gttcacaatg
     2401 aaaccaagaa tgaattggag aagatgttga aatgtaatga ggagcaccca gcttatcttg
     2461 caagtgatga aataaccaca gtccggaaga accttgaatc ccgaggagta gaagtagatc
     2521 caagcttgat taaggatact tggcatcaag tttatagaag acatttttta aaaacagctc
     2581 taaaccattg taacctttgt cgaagaggtt tttattacta ccaaaggcat tttgtagatt
     2641 ctgagttgga atgcaatgat gtggtcttgt tttggcgtat acagcgcatg cttgctatca
     2701 ccgcaaatac tttaaggcaa caacttacaa atactgaagt taggcgatta gagaaaaatg
     2761 ttaaagaggt attggaagat tttgctgaag atggtgagaa gaagattaaa ttgcttactg
     2821 gtaaacgcgt tcaactggcg gaagacctca agaaagttag agaaattcaa gaaaaacttg
     2881 atgctttcat tgaagctctt catcaggaga aagggctagc gctaccggtc gccaccatgg
     2941 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3001 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3061 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3121 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3181 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3241 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3301 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3361 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3421 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3481 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3541 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3601 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3661 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3721 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3901 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4441 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4501 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4681 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4741 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4801 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4861 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4921 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4981 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5041 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5101 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5161 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5221 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5281 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5341 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5401 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5461 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5521 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5581 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5641 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5701 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5761 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5821 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5881 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5941 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6001 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6061 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6121 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6241 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6421 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6961 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     7021 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7081 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7141 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7201 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     7261 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     7321 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     7381 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     7441 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     7501 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     7561 gt
//
