LOCUS       sequence-404953-        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: Palpha2
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2951..2960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2957..2959,2960..2962,2963..3676)
                     /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(3002..3023)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3263..3282)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3610..3631
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3890..3909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3927..4382)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4064..4085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4276..4295
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4409..4513
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4515..4872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4515..4706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4538..4558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4723..4858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4907..5701
                     /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             4907..5701
                     /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(4961..4980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5571..5590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5889..5908)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5933..5980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7072..7375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7376..7562,1..17)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7530..7550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cgatgtggcg
       61 actacgtcgg gccgctgtgg cctgtgaggt ctgccagtct ttagtgaaac acagctctgg
      121 aataaaagga agtttaccac tacaaaaact acatctggtt tcacgaagca tttatcattc
      181 acatcatcct accttaaagc ttcaacgacc ccaattaagg acatcctttc agcagttctc
      241 ttctctgaca aaccttcctt tacgtaaact gaaattctct ccaattaaat atggctacca
      301 gcctcgcagg aatttttggc cagcaagatt agctacgaga ctcttaaaac ttcgctatct
      361 catactagga tcggctgttg ggggtggcta cacagccaaa aagacttttg atcagtggaa
      421 agatatgata ccggacctta gtgaatataa atggattgtg cctgacattg tgtgggaaat
      481 tgatgagtat atcgattttg agaaaattag aaaagccctt cctaattcag aagaccttgt
      541 aaagttagca ccagactttg acaagattgt tgaaagcctt agcttattga aggacttttt
      601 tacctcaggt tctccggaag aaacggcgtt tagagcaaca gatcgtggat ctgaaagtga
      661 caagcatttt agaaaggtgt cagacaaaga gaaaattgac caacttcagg aagaacttct
      721 gcacactcag ttgaagtatc agagaatctt ggaacgatta gaaaaggaga acaaagaatt
      781 gagaaaatta gtattgcaga aagatgacaa aggcattcat catagaaagc ttaagaaatc
      841 tttgattgac atgtattctg aagttcttga tgttctctct gattatgatg ccagttataa
      901 tacgcaagat catctgccac gggttgttgt ggttggagat cagagtgctg gaaagactag
      961 tgtgttggaa atgattgccc aagctcgaat attcccaaga ggatctgggg agatgatgac
     1021 acgttctcca gttaaggtga ctctgagtga aggtcctcac catgtggccc tatttaaaga
     1081 tagttctcgg gagtttgatc ttaccaaaga agaagatctt gcagcattaa gacatgaaat
     1141 agaacttcga atgaggaaaa atgtgaaaga aggctgtacc gttagccctg agaccatatc
     1201 cttaaatgta aaaggccctg gactacagag gatggtgctt gttgacttac caggtgtgat
     1261 taatactgtg acatcaggca tggctcctga cacaaaggaa actattttca gtatcagcaa
     1321 agcttacatg cagaatccta atgccatcat actgtgtatt caagatggat ctgtggatgc
     1381 tgaacgcagt attgttacag acttggtcag tcaaatggac cctcatggaa ggagaaccat
     1441 attcgttttg accaaagtag acctggcaga gaaaaatgta gccagtccaa gcaggattca
     1501 gcagataatt gaaggaaagc tcttcccaat gaaagcttta ggttattttg ctgttgtaac
     1561 aggaaaaggg aacagctctg aaagcattga agctataaga gaatatgaag aagagttttt
     1621 tcagaattca aagctcctaa agacaagcat gctaaaggca caccaagtga ctacaagaaa
     1681 tttaagcctt gcagtatcag actgcttttg gaaaatggta cgagagtctg ttgaacaaca
     1741 ggctgatagt ttcaaagcaa cacgttttaa ccttgaaact gaatggaaga ataactatcc
     1801 tcgcctgcgg gaacttgacc ggaatgaact atttgaaaaa gctaaaaatg aaatccttga
     1861 tgaagttatc agtctgagcc aggttacacc aaaacattgg gaggaaatcc ttcaacaatc
     1921 tttgtgggaa agagtatcaa ctcatgtgat tgaaaacatc taccttccag ctgcgcagac
     1981 catgaattca ggaactttta acaccacagt ggatatcaag cttaaacagt ggactgataa
     2041 acaacttcct aataaagcag tagaggttgc ttgggagacc ctacaagaag aattttcccg
     2101 ctttatgaca gaaccgaaag ggaaagagca tgatgacata tttgataaac ttaaagaggc
     2161 cgttaaggaa gaaagtatta aacgacacaa gtggaatgac tttgcggagg acagcttgag
     2221 ggttattcaa cacaatgctt tggaagaccg atccatatct gataaacagc aatgggatgc
     2281 agctatttat tttatggaag aggctctgca ggctcgtctc aaggatactg aaaatgcaat
     2341 tgaaaacatg gtgggtccag acgcggcagc ggcagctgca gccgcggcgg ctgcgaccca
     2401 agaacagtgt gttcacaatg aaaccaagaa tgaattggag aagatgttga aatgtaatga
     2461 ggagcaccca gcttatcttg caagtgatga aataaccaca gtccggaaga accttgaatc
     2521 ccgaggagta gaagtagatc caagcttgat taaggatact tggcatcaag tttatagaag
     2581 acatttttta aaaacagctc taaaccattg taacctttgt cgaagaggtt tttattacta
     2641 ccaaaggcat tttgtagatt ctgagttgga atgcaatgat gtggtcttgt tttggcgtat
     2701 acagcgcatg cttgctatca ccgcaaatac tttaaggcaa caacttacaa atactgaagt
     2761 taggcgatta gagaaaaatg ttaaagaggt attggaagat tttgctgaag atggtgagaa
     2821 gaagattaaa ttgcttactg gtaaacgcgt tcaactggcg gaagacctca agaaagttag
     2881 agaaattcaa gaaaaacttg atgctttcat tgaagctctt catcaggaga aagggctagc
     2941 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3001 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3061 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3121 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3181 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3241 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3301 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3361 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3421 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3481 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3541 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     3601 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3661 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3721 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3781 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3901 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3961 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4021 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4081 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4141 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4201 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4261 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4321 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4381 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4441 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4501 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4741 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4801 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4861 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4921 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4981 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5041 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5101 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5161 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5221 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5281 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5341 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5401 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5461 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5521 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5581 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5641 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5701 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5761 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5821 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5881 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5941 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6001 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     6061 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6121 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     7021 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7081 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7141 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7201 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7261 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7321 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     7381 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7441 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7501 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7561 gg
//
