LOCUS       sequence-404947-        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-5
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2949..2958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2955..2957,2958..2960,2961..3674)
                     /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(3000..3021)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3261..3280)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3608..3629
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3797..3918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3834..3853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3925..4380)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4083)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4274..4293
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4407..4511
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4513..4870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4513..4704
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4536..4556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4721..4856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4905..5699
                     /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             4905..5699
                     /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(4959..4978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5569..5588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5887..5906)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5931..5978
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6307..6895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6796..6815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7070..7373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7374..7562,1..15)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7528..7548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg atgtggcgac
       61 tacgtcgggc cgctgtggcc tgtgaggtct gccagtcttt agtgaaacac agctctggaa
      121 taaaaggaag tttaccacta caaaaactac atctggtttc acgaagcatt tatcattcac
      181 atcatcctac cttaaagctt caacgacccc aattaaggac atcctttcag cagttctctt
      241 ctctgacaaa ccttccttta cgtaaactga aattctctcc aattaaatat ggctaccagc
      301 ctcgcaggaa tttttggcca gcaagattag ctacgagact cttaaaactt cgctatctca
      361 tactaggatc ggctgttggg ggtggctaca cagccaaaaa gacttttgat cagtggaaag
      421 atatgatacc ggaccttagt gaatataaat ggattgtgcc tgacattgtg tgggaaattg
      481 atgagtatat cgattttgag aaaattagaa aagcccttcc taattcagaa gaccttgtaa
      541 agttagcacc agactttgac aagattgttg aaagccttag cttattgaag gactttttta
      601 cctcaggttc tccggaagaa acggcgttta gagcaacaga tcgtggatct gaaagtgaca
      661 agcattttag aaaggtgtca gacaaagaga aaattgacca acttcaggaa gaacttctgc
      721 acactcagtt gaagtatcag agaatcttgg aacgattaga aaaggagaac aaagaattga
      781 gaaaattagt attgcagaaa gatgacaaag gcattcatca tagaaagctt aagaaatctt
      841 tgattgacat gtattctgaa gttcttgatg ttctctctga ttatgatgcc agttataata
      901 cgcaagatca tctgccacgg gttgttgtgg ttggagatca gagtgctgga aagactagtg
      961 tgttggaaat gattgcccaa gctcgaatat tcccaagagg atctggggag atgatgacac
     1021 gttctccagt taaggtgact ctgagtgaag gtcctcacca tgtggcccta tttaaagata
     1081 gttctcggga gtttgatctt accaaagaag aagatcttgc agcattaaga catgaaatag
     1141 aacttcgaat gaggaaaaat gtgaaagaag gctgtaccgt tagccctgag accatatcct
     1201 taaatgtaaa aggccctgga ctacagagga tggtgcttgt tgacttacca ggtgtgatta
     1261 atactgtgac atcaggcatg gctcctgaca caaaggaaac tattttcagt atcagcaaag
     1321 cttacatgca gaatcctaat gccatcatac tgtgtattca agatggatct gtggatgctg
     1381 cgcgcagtat tgttacagac ttggtcagtc aaatggaccc tcatggaagg agaaccatat
     1441 tcgttttgac caaagtagac ctggcagaga aaaatgtagc cagtccaagc aggattcagc
     1501 agataattga aggaaagctc ttcccaatga aagctttagg ttattttgct gttgtaacag
     1561 gaaaagggaa cagctctgaa agcattgaag ctataagaga atatgaagaa gagttttttc
     1621 agaattcaaa gctcctaaag acaagcatgc taaaggcaca ccaagtgact acaagaaatt
     1681 taagccttgc agtatcagac tgcttttgga aaatggtacg agagtctgtt gaacaacagg
     1741 ctgatagttt caaagcaaca cgttttaacc ttgaaactga atggaagaat aactatcctc
     1801 gcctgcggga acttgaccgg aatgaactat ttgaaaaagc taaaaatgaa atccttgatg
     1861 aagttatcag tctgagccag gttacaccaa aacattggga ggaaatcctt caacaatctt
     1921 tgtgggaaag agtatcaact catgtgattg aaaacatcta ccttccagct gcgcagacca
     1981 tgaattcagg aacttttaac accacagtgg atatcaagct taaacagtgg actgataaac
     2041 aacttcctaa taaagcagta gaggttgctt gggagaccct acaagaagaa ttttcccgct
     2101 ttatgacaga accgaaaggg aaagagcatg atgacatatt tgataaactt aaagaggccg
     2161 ttaaggaaga aagtattaaa cgacacaagt ggaatgactt tgcggaggac agcttgaggg
     2221 ttattcaaca caatgctttg gaagaccgat ccatatctga taaacagcaa tgggatgcag
     2281 ctatttattt tatggaagag gctctgcagg ctcgtctcaa ggatactgaa aatgcaattg
     2341 aaaacatggt gggtccagac tggaaaaaga ggtggttata ctggaagaat cggacccaag
     2401 aacagtgtgt tcacaatgaa accaagaatg aattggagaa gatgttgaaa tgtaatgagg
     2461 agcacccagc ttatcttgca agtgatgaaa taaccacagt ccggaagaac cttgaatccc
     2521 gaggagtaga agtagatcca agcttgatta aggatacttg gcatcaagtt tatagaagac
     2581 attttttaaa aacagctcta aaccattgta acctttgtcg aagaggtttt tattactacc
     2641 aaaggcattt tgtagattct gagttggaat gcaatgatgt ggtcttgttt tggcgtatac
     2701 agcgcatgct tgctatcacc gcaaatactt taaggcaaca acttacaaat actgaagtta
     2761 ggcgattaga gaaaaatgtt aaagaggtat tggaagattt tgctgaagat ggtgagaaga
     2821 agattaaatt gcttactggt aaacgcgttc aactggcgga agacctcaag aaagttagag
     2881 aaattcaaga aaaacttgat gctttcattg aagctcttca tcaggagaaa gggctagcgc
     2941 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3001 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtcaggc gagggcgagg
     3061 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     3121 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3181 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3241 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3301 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3361 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     3421 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3481 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3541 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     3601 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3661 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     3721 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3781 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtacaa
     3901 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3961 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4021 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4081 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4141 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4201 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4261 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4321 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4501 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4621 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4681 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4801 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4861 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4921 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4981 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5041 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5101 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5161 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5221 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5281 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5341 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5401 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5461 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5521 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5581 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5641 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5701 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5761 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5821 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5881 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5941 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6001 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6061 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6121 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6421 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6961 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     7021 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7081 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     7141 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     7201 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     7261 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     7321 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     7381 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     7441 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     7501 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     7561 tc
//
